Giving Evolutionary Biologists the Finger!

Evolution of KRAB Zinc Finger Proteins vs. the Law of Large Numbers

There are patterns in biology that violate the law of large numbers, and thus suggests Intelligent Design or at the very least statistical miracles. The pattern involves KRAB-ZnF proteins that have multiple zinc finger domains side by side that are inexact copies of each other and would require a scenario of co-evolution of their DNA binding partners with every additional zinc-finger insertion — a scenario indistinguishable from a miracle.

The role of a zinc finger can be something like a clamp or a lock. Each zinc finger in a KRAB-ZnF protein is fine-tuned to connect with a DNA much like a lock (the zinc finger) can receive a key (like DNA). Here is a depiction of a KRAB-ZnF protein with 4 zinc fingers as part of a large chromatin modifying complex. The four zinc fingers are marked “ZN” and connect to DNA:

https://journals.plos.org/plosone/article/figure/image?id=10.1371/journal.pone.0023747.g005&size=large

The evolutionary explanation of side-by-side repeated patterns of zinc fingers such as in KRAB-ZnF proteins shows a severe lack of critical thinking by evolutionary biologists who pretend “phylogenetic methods” are adequate explanations of mechanical feasibility of common descent.

To illustrate the problem, consider the KRAB-ZnF protein known as ZNF136. For reference, this is the amino acid sequence of ZNF136:
https://www.uniprot.org/uniprot/P52737.fasta

This is an amino acid fragment contained within the ZNF136 protein

TGEKLYDCKECGKTFFSLKRIRRHIITH

This short sequence is called a zinc finger domain which in 3D looks like this:

https://sciencescienceeverywhere.files.wordpress.com/2015/08/figure-11.jpg

Wiki gives a description of the function of zinc fingers in proteins that have them:

http://wikipedia.org/wiki/Zinc_finger

A zinc finger domain requires two “C” amino acids and two “H” amino acids placed in the right positions. It requires a few other things too…

There are 13 zinc fingers in the ZNF136 protein and these are their sequences:

TGEKLYDCKECGKTFFSLKRIRRHIITH

SGYTPYKCKVCGKAFDYPSRFRTHERSH

TGEKPYECQECGKAFTCITSVRRHMIKH

TGDGPYKCKVCGKPFHSLSSFQVHERIH

TGEKPFKCKQCGKAFSCSPTLRIHERTH

TGEKPYECKQCGKAFSYLPSLRLHERIH

TGEKPFVCKQCGKAFRSASTFQIHERTH

TGEKPYECKECGEAFSCIPSMRRHMIKH

TGEGPYKCKVCGKPFHSLSPFRIHERTH

TGEKPYVCKHCGKAFVSSTSIRIHERTH

TGEKPYECKQCGKAFSYLNSFRTHEMIH

TGEKPFECKRCGKAFRSSSSFRLHERTH

TGQKPYHCKECGKAYSCRASFQRHMLTH

For completeness, there is a degenerate zinc finger in ZNF136 with the sequence “YGEKPDTRNQCWKPFSSHHSFRTHEIIH”

Why are the Zinc Fingers so different in sequence (except for the conserved amino acids)? To target a section of DNA, the zinc finger must be tuned to target it. Think of the zinc finger like a lock and DNA as a key that fits into the lock. In fact, for both the study of biology and medical applications, humans have a desire to make their own zinc fingers — like lock smiths. To bind a large segments of DNA, side-by-side zinc-fingers have to be tuned to their respective side-by-side DNA partners such as illustrated here:

https://journals.plos.org/plosone/article/figure/image?id=10.1371/journal.pone.0023747.g005&size=large

By the way, there is a website that helps researchers construct the right amino acid sequence to make a zinc finger for a particular DNA target:

https://www.scripps.edu/barbas/zfdesign/zfdesignhome.php

Now, to visualize the critical/conserved amino acids, see the protein sequence here with highlights on “C” and “H” amino acids.

http://theskepticalzone.com/wp/wp-content/uploads/2019/04/znf136_zfC2H21.png

Note one of the lines is not exactly like the other lines in that it is missing a “C”. This is the degenerate zinc finger mentioned above. So there is 1 degenerate zinc finger and 13 functional ones.

From this diagram it is apparent that the regular appearance of “C” and “H” is a violation of the law of large numbers, hence this pattern is not due to random point mutation alone. To “solve” this problem, Darwinist explain the pattern through segment duplication followed by some point mutation and natural selection, but this is not mechanically feasible either!

To understand why, let the reader first ponder the alignment I made of the Zinc Fingers in the ZNF136 protein using MEGA 6.0/MUSCLE software:

http://theskepticalzone.com/wp/wp-content/uploads/2019/04/znf136_zfC2H2_muscle1.png

Then let the reader, consider the distance matrix generated by MEGA 6.0 which measures the number of nucleotide and percent differences between the zinc fingers.

http://www.creationevolutionuniversity.org/public_blogs/reddit/znf136_distance_matrix.xls

(All of the above results are reproducible, so I leave it to interested parties wanting to confirm the results to do so.)

For the duplication to work, at a bare minimum the right 84 nucleotide segment must be chosen, and then perfectly positioned for insertion so as not to break a pre-existing zinc finger. But supposing the duplication succeeds, why are the zinc finger’s conserved features involving “C” and “H” and other amino acids preserved and not eventually erased by point mutation given we obviously see the zinc fingers are different from each other. To preserve the “C” and “H” and other necessary amino acids in a zinc finger, the new zinc finger needs to be under selection. But in that case one is simply concocting a “just so” story for those newly minted zinc fingers without any respect for the difficulty of such a “just so” story being probable — and it is not probable!

To understand the problem of such a “just so” story, recall zinc fingers bind to DNA regions. Btw, this includes DNA regions such as ERVs! And ERVs are indicated to participate in the Stem Cell Pluripotency regulatory network:

https://www.nature.com/articles/nature13804

Further, the KRAB-ZnF protein is part of an incredibly complex machine that does chromatin modification (as shown above) by often attaching to ERV targets. But this would require that ERVs (or whatever DNA target) needs to co-evolve with the KRAB-ZnFs that attach to them!

https://genome.cshlp.org/content/21/11/1800?ijkey=f1f42a3747733fd430f8966c8014affc0af066e8&keytype2=tf_ipsecsha

Do these researchers even consider the fact such evolution would have to be instantaneous otherwise it would degrade function and not let the genome have the opportunity to adapt to the new accidental copy of the zinc finger because the duplication would immediately be selected against!

The above KRAB-ZnF complex is like a read/write head acting on Chromatin. Chromatin itself is an amazing mind-boggling design akin to computer ROM and RAM in one.

Again, the difficulty of evolution via random insertion/duplication mutations followed by point mutations is that such events would disrupt the binding of an already operational set of zinc fingers. For example, suppose we have an array of 10 zinc fingers side-by-side that collectively bind to a target DNA. Suppose one zinc-finger is duplicated and the number of zinc fingers is increased from 10 to 11. Oh well, the binding ability is broken or at best compromised, much like adding a single letter to a pre-existing password!

In sum, there is a violation of the law of large numbers in KRAB-ZnF proteins which is not explained by random mutation, nor random segment duplication followed by some point mutation and fixed by natural selection. Some other mechanism for the emergence of such proteins is indicated and would likely be indistinguishable from a miracle. Given the importance of such zinc finger proteins in the control of ERVs which are important in the stem cell pluripotency regulatory circuits, the origin of KRAB-ZnFs is even more miraculous.

476 thoughts on “Giving Evolutionary Biologists the Finger!

  1. colewd: Why don’t you and Entropy start by demonstrating you understand Sal’s arguments. You guys are talking over each other. You as a moderator need to be a leader and right now you are leading this site right off a cliff.

    I am curious Bill, do you honestly think that there is an equivalency to the way in which Sal and the rest of us are “talking over each other”, or do you know that phrasing it that way is a handy rhetorical gambit to make the two sides appear equivalent, in a “Trump on Charlottesville” kinda way?

    Here’s Sal’s argument:

    Sal notes that there are many Zinc finger proteins which bind to many different DNA sequences.
    Each protein-target pair is an example of irreducible complexity, since the protein consists of a number of repeats, each with lock-and-key-like specificity for the corresponding DNA sequence, just like tumblers in a combination lock.
    It is implausibly unlikely that one such 13 finger protein could co-evolve with its corresponding target DNA sequence. Each extra finger would have to arise simultaneously with the corresponding DNA target.
    Based on simple statistics, the LLN, we can demonstrate that the probability of random mutations creating these sequences is vanishingly small, since the repeated distribution of the C’s and H’s in the fingers is waaay beyond the expected random distribution.
    Evilutionists claim that these fingers could have arisen via duplication. There are a number of problems with this: how does the cell know which bits of DNA to duplicate? How is it that the Zinc fingers are the bits that are duplicated? How is it that the duplication is PRECISELY 84 nucleotides for each of the 13 separate duplications? The probability of this happening is vanishingly small. Chances are, one of the extra copies will be out of frame, then BOOM! No Zinc Finger protein!

    Ack! My brain hurts.

  2. colewd

    Does anyone have experimental evidence that these sequences have flexibility? Are they highly preserved?.

    I suggest you read the OP, and the links therein. Especially the bits about creating DNA binding domains to order.

  3. DNA_Jock:

    failing to notice that “degenerate” repeat 1 isn’t even a full Zinc finger

    From the Opening Post

    stcordova:

    Note one of the lines is not exactly like the other lines in that it is missing a “C”. This is the degenerate zinc finger mentioned above.

    Is that the best you got? LOL! Just make crap up even when I said otherwise from the very start. Thanks for the entertainment.

    Bill, thanks for showing. It’s not worth paying attention to their drivel. That’s example right there.

  4. This is a teachable moment to see how the moderator of TSZ makes up crap:

    DNA_Jock says falsely of me:

    failing to notice that “degenerate” repeat 1 isn’t even a full Zinc finger

    but in contrast I said from the very opening the degnerate finger wasn’t functional.

    stcordova said at the opening:

    Note one of the lines is not exactly like the other lines in that it is missing a “C”. This is the degenerate zinc finger mentioned above. So there is 1 degenerate zinc finger and 13 functional ones.

    Will he make a retraction now that’s been called out on attributing to me a claim I didn’t make?

    Bill, don’t be bamboozled by this sort of crap. I may not take the time to call out all of it since it’s going to take a lot of time.

    Don’t worry about defending me. I’m not here to advertise my work, I’m here to get editorial review and find out what sort of falsehoods, misrepresentations, and other crap my critics might throw at it so I can better compose my essays.

  5. What I wrote.

    Given the opportunity to demonstrate his comprehension, Sal whines an irrelevance about the rooting of the tree (perhaps failing to notice that “degenerate” repeat 1 isn’t even a full Zinc finger) and waffles some utter rubbish about…

    My apologies, Sal. You did in fact notice that repeat 1 is not a full Zinc finger.
    Now that we have that out of the way, perhaps you will ease up on the misrepresentation and address the remainder of everyone’s demonstrations of your errors.
    Or not. Your choice.

  6. A teachable moment:

    This is how to criticize an argument when you can’t refute the actual points. Criticize something that wasn’t ever claimed or said. Here is an example:

    DNA_Jock says falsely of me:

    failing to notice that “degenerate” repeat 1 isn’t even a full Zinc finger

    Ah but DNA_jock seems to have the audacity to say stuff like that given I said from the beginning:

    http://wikipedia.org/wiki/Zinc_finger

    A zinc finger domain requires two “C” amino acids and two “H” amino acids placed in the right positions. It requires a few other things too…

    Note one of the lines is not exactly like the other lines in that it is missing a “C”. This is the degenerate zinc finger mentioned above. So there is 1 degenerate zinc finger and 13 functional ones.

    To say:

    So there is 1 degenerate zinc finger and 13 functional ones.

    Means the degenerate one is NOT functional.

    But this doesn’t stop DNA_Jock from saying:

    DNA_Jock:

    DNA_Jock says falsely of me:

    failing to notice that “degenerate” repeat 1 isn’t even a full Zinc finger

    Will he make a retraction? Any bets, or will he perpetuate the falsehood, pretend he wasn’t called out on it.

    The other question is whether it’s worth my time if this is the best he’s got because he’s certainly not refuted the other problems I pointed out.

    As far as the supposed clustering, the unrooted tree does a better job of identifying clusters, I didn’t have to notice or not notice, I let the computation made by evolutionary biologists do the talking as in:

  7. DNA_Jock:

    The dispositive thing about this tree is that repeat #4 is most closely related to #9, 5 and 10,

    Only if you root the tree like Evograd did.

    The unrooted tree which isn’t as prejudiced, and the one I did, gave a different clustering.

    There was a typo above where I used the #13 twice, namely

    #13 499-526,
    #13 471-442

    when it should have been
    #13 471-498
    #14 499-526

    But other than that, both graphs I generated refute DNA_Jock’s claim, whether #13 (above) or #14(corrected), the clustering isn’t as DNA_Jock says UNLESS one forces a rooting like evograd did which is imposing something of a pre-conceived phylogeny to begin with, and in the absence of other data,
    that is circular reasoning.

    The Neighbor Joining does a different clustering:

    2, 4, 9,14

    But if we drop neighbor joining, the actual distances from 4 are:

    1 vs. 4 = 16
    2 vs 4 = 12
    3 vs 4 = 16
    4 vs 4 = 0
    5 vs 4 = 16
    6 vs 4 = 13
    7 vs 4 = 11
    8 vs 4 = 15
    9 vs 4 = 5
    10 vs 4 = 15
    11 vs 4 = 11
    12 vs 4 = 12
    13 vs 4 = 11

    Moral: nearness is in the eye of the beholder and IT’S FREAKING IRRELEVANT TO THE PROBLEMS THAT I’VE POINTED OUT.

    One can assume one phylogeny exists, it doesn’t refute the point that this is an improbable structure for the reasons stated.

  8. DNA_Jock:
    What I wrote.

    My apologies, Sal. You did in fact notice that repeat 1 is not a full Zinc finger.
    Now that we have that out of the way, perhaps you will ease up on the misrepresentation and address the remainder of everyone’s demonstrations of your errors.
    Or not. Your choice.

    Are you going to retract you howler that adding a zinc finger copy increases binding affinity for a PRE-EXISTING cis-regulatory element? 🙂

  9. stcordova: Are you going to retract you howler that adding a zinc finger copy increases binding affinity for a PRE-EXISTING cis-regulatory element? 🙂

    Do explain.
    Really. This will be fun.
    I’m all ears.

  10. DNA_Jock: Do explain.
    Really. This will be fun.
    I’m all ears.

    DNA_Jock claims making tandem IDENTICAL repeats increases binding affinity.

    Well I was speaking of binding affinity to a PRE-EXISTING DNA target RELATIVE to the affinity the tandem repeat mutant will bind to.

    I wasn’t speaking about some OTHER concoction where it might actually increase binding affinity. I was talking zinc-finger-to-DNA binding, as in a zinc-finger to DNA cis-regulatory element of a gene.

    From:

    http://zifit.partners.org/ZiFiT_v3.3/Program_use.aspx#_ZiFiT_ZF_Array

    Multiple ZFs can be linked together to recognize a specific and preferably unique sequence in double-stranded genomic DNA. When multiple ZFs are joined together, the resulting multi-finger array recognizes a longer target DNA sequence. Multi-finger arrays can be fused to and thereby target transcriptional activation domains, repressors, or nucleases to specific locations in the genome.

    So the finger binding affinities are:

    ZF3 = GCG
    ZF2 = TGG
    ZF1 = GCG

    That’s a gross simplification because actually each ZF binds up to 4 bases and must over lap in binding with the adjacent ZF as mentioned above! But I’ll go with the simple diagram for now:

    But lets suppose we had tandem repeat zinc fingers binding to the same motif : GCG

    ZF3 = GCG
    ZF2 = GCG
    ZF1 = GCG

    So the composite target of identical tandem repeats is:

    GCGGCGGCG

    not

    GCGTGGGCG

    Now suppose the original array was only 1 zinc finger that bound to GCG. Well that’s awfully non-specific and would have binding affinity to lots of places on the genome by random chance! But now lets add 2 tandem repeats (using the simplified logic which isn’t exactly correct because of the 4bp and overlap problem mentioned earlier by me). The binding target is now:

    GCGGCGCGC

    thus when the protein having only 1 zinc finger might previously have some chance of binding to

    TTTGCGAAA

    it will NOT increase the likelihood that the array of 3 tandem zinc fingers will bind to that relative to this target:

    GCGGCGCGC

    One might argue that it will could still bind to the old target of:

    TTTGCGAAA

    But if we include the problem of overlap mentioned above where non-symetric 4base DNA sequences are targets of the zinc finger, then increased binding is dubious.

    What do I mean? The actual situation is the ZF can target 4 bases like:

    ACGA

    So a tandem 2 repeat zinc finger could bind this:

    ACGACGA

    where
    ZF1 binds ACGA
    ZF2 binds ACGA

    ACGA has some symmetry between the A at the start and A at the end. But consider a non symmetric target for the original zinc finger:

    ACGT

    So a tandem repeat will have the following:

    ZF1: ACGT
    ZF2: ACGT

    The composite recognition sequence would be:

    ACG?ACG

    where question mark is for how the 2 identical repeat tandem will handle the overlapping position.

  11. Since DNA_jock apologized for falsely saying I failed to point out the degenerate zinc finger is non-functional, I will extend a little bit of charity.

    This stuff is confusing, and it is possible DNA_Jock mis-understood what I said. I will attempt to clarify. If a solitary zinc finger targets this motif:

    ACGA

    then a partial list of DNA segments the zinc finger could park in the vicinity of could be (with the ZF binding target in caps, and the flanking DNA in lower case):

    1 ACGA aaaa

    2 ACGA aaac

    3 ACGA aaag

    4 ACGA aaat

    5 ACGA aaca

    6 ACGA aacc

    7 ACGA aacg

    8 ACGA aact

    9 ACGA aaga

    10 ACGA aagc

    11 ACGA aagg

    12 ACGA aagt

    13 ACGA aata

    14 ACGA aatc

    15 ACGA aatg

    16 ACGA aatt

    16 ACGA aatt

    17 ACGA acaa

    18 ACGA acac

    19 ACGA acag

    20 ACGA acat

    21 ACGA acca

    22 ACGA accc

    23 ACGA accg

    24 ACGA acct

    25 ACGA acga

    ……

    64 ACGA tttt

    Ok so now let’s say we have that tandem repeat described above what will the affinity be for?

    ACGACGA

    Well ACGACGA is #25 on the list above

    So this sequence with have a binding affinity greater than its sisters in the above list. Hence the previous 63 other binding targets have reduced likelihood of being bound RELATIVE to sequnce

    #25 ACGACGA

    in that sense the other 63 sequences have a lessened affinity relative to the sequence that has the strongest affinity to the new tandem repeat.

    That hopefully clarifies what I meant by reduced affinity. Since that verbage on my part may have caused confusion, I’ll think on how to phrase it differently.

    But the fundamental point remains that a former potential function is now altered and even in this case, assuming HYPOTHETICALLY it was functional to target ACGA with one zinc finger, now 63 prior DNA sites have been rendered functionless by a “random tandem” repeat of a pre-existing zinc finger.

    That’s the BAD JUJU I was talking about.

  12. Inappropriately, I’m reminded of the Charge of the Light Brigade – both the poem and the event.

  13. If you had a duplication of a protein segment with affinity for a particular DNA sequence, I really don’t get how you’d get reduced binding as a result. Some of the protein molecules would bind using site A, some site B. The two sites aren’t competing with each other within any given molecule. Displacement of a B-bound molecule by an A would be no different from an A shoving another A out the way. Detachment of a single molecule at B and re-attachment at A is still bound.

    When bound by site A, there will be some interactions of B with flanking regions. And vice versa. This would often tend to increase the overall binding energy … wouldn’t It?

  14. Allan Miller: When bound by site A, there will be some interactions of B with flanking regions. And vice versa. This would often tend to increase the overall binding energy … wouldn’t It?

    Yep. Imagine a piece of tape that you stick to something. One of the flanking sides of the tape has a bit of hair on it so it doesn’t stick as well as the rest, but it still sticks.

  15. Allan Miller:
    If you had a duplication of a protein segment with affinity for a particular DNA sequence, I really don’t get how you’d get reduced binding as a result. Some of the protein molecules would bind using site A, some site B. The two sites aren’t competing with each other within any given molecule. Displacement of a B-bound molecule by an A would be no different from an A shoving another A out the way. Detachment of a single molecule at B and re-attachment at A is still bound.

    When bound by site A, there will be some interactions of B with flanking regions. And vice versa. This would often tend to increase the overall binding energy … wouldn’t It?

    I was speaking in terms of RELATIVE binding affinities rather than absolute affinities.

    In my preceding comment the binding affinity in my pedagogical/hypothetical model for the 64 sites was the same.

    The affinity is 1/KD where KD is the dissociation constant.

    Prior to the tandem repeat event

    1/KD_1 = 1/KD_2 = …. 1/KD_25 .. 1/KD_64

    after the tandem repeat event

    1/KD_25 > 1/KD_1, 1/KD_2….1/KD_24, 1/KD26…1/KD_64

    That’s what’s important. If this were not substantial enough of a difference, then we might expect the multi-zinc finger arrays to have their individual zinc fingers bind all over the place, but that apparently doesn’t happen — certainly the ZFIT consortium, when they intelligently design a zinc finger array for their custormers, they expect it to bind to the specified target, not have the individual zinc fingers randomly attach all over a genome!

  16. stcordova: But the fundamental point remains that a former potential function is now altered and even in this case, assuming HYPOTHETICALLY it was functional to target ACGA with one zinc finger, now 63 prior DNA sites have been rendered functionless by a “random tandem” repeat of a pre-existing zinc finger.

    But you don’t know that, you’re just asserting it would be rendered non functional. There’s just no reason to think that.

  17. stcordova: I was speaking in terms of RELATIVE binding affinities rather than absolute affinities.

    Relative to what? The singleton’s affinity? It can easily go up. It would be perverse to deny this.

    [frantic Googling reveals…]

    The affinity is 1/KD where KD is the dissociation constant …

    You can’t decompose it in that per-site way. Two molecules have an overall affinity. It’s that which determines how well they stick together. Obviously, the bits and pieces of the molecules contribute to that overall affinity, by the electrostatic zone of influence of each bit of zwitterion or whatever, but it’s nonsensical to try and separate out the contribution of each DNA base.

    If a single zinc finger targets a particular set of bases, why would a dimer go anywhere else?

  18. The peculiar notion being peddled here is that, if a single finger targets a given sequence, a dimer will helplessly settle halfway between the two, in a perpetual – but weak! – arm-wrestle for the target. Before heading off and binding somewhere else instead, for which there is no affinity.

  19. Allan Miller: Relative to what? The singleton’s affinity? It can easily go up. It would be perverse to deny this.

    You can’t decompose it in that per-site way. Two molecules have an overall affinity. It’s that which determines how well they stick together. Obviously, the bits and pieces of the molecules contribute to that overall affinity, by the electrostatic zone of influence of each bit of zwitterion or whatever, but it’s nonsensical to try and separate out the contribution of each DNA base.

    If a single zinc finger targets a particular set of bases, why would a dimer go anywhere else?

    For convenience I will call 1/KD = KA

    Do you think KA_25 > KA_1, KA_2, … ???

    But let’s say a random tandem repeat event doesn’t make KA_25 THE chosen one, but rather KA_X

    So in principle KA_X > KA_1, KA_2 ……KA_64

    So even in that case my point is proven. Previously cis-regulatory targets are now not as likely to be bound as before. QED.

  20. stcordova: So even in that case my point is proven. Previously cis-regulatory targets are now not as likely to be bound as before. QED.

    What the fuck are you talking about?

  21. Rumraket:
    What the fuck are you talking about?

    He’s saying that because he refuses, and or lacks the background/abilities, to even try and understand explanations to the contrary, he’s right.

  22. stcordova,

    Not sure you’re getting my point. If a single finger binds well to a particular sequence, why would the binding of a dimer be less strong? I mean, there may be steric interference or electrostatic effects between the monomers diminishing the substrate interaction, but that aside, to the extent that subunit B is shoving A out the way, or vice versa, they are doing so because of their mutual attraction to the single substrate.

  23. Allan:

    f a single finger binds well to a particular sequence, why would the binding of a dimer be less strong?

    The dimer binding is stronger than the monomer. That was the case in #25 ACGACGA above, or equivalently the dissociation is less for #25.

    For that reason, with the same amount of the transciption factor concentration, more of the transcription will bind to #25 and less to the other 63 cis regulatory regions in my pedagogical example.

  24. stcordova,

    For that reason, with the same amount of the transciption factor concentration, more of the transcription will bind to #25 and less to the other 63 cis regulatory regions in my pedagogical example.

    What will happen given this condition?

  25. colewd:
    stcordova,

    What will happen given this condition?

    IF the other 63 sites were important, and now they are getting serviced less or not at all, in general this is the result:

  26. stcordova,

    IF the other 63 sites were important, and now they are getting serviced less or not at all, in general this is the result:

    This can certainly be true with transcription factors. There is a range of acceptable function or transcription rates however.

  27. See, Rumraket, I told you that that was his point.
    Do you want to tell him, or shall I, just how much of the protein will be bound to sequences that are completely unrelated to any target sequences?
    Kudos to colewd for his “I don’t think that will matter” response.

  28. DNA_Jock: See, Rumraket, I told you that that was his point.

    Yeah Rumraket. You tell him right Jock, you said so yourself:

    Giving Evolutionary Biologists the Finger!

    The tandem finger would bind tighter and more selectively

    DNA_Jock even bolded it like that. Yeah. MORE SELECTIVELY. As in tighter to the new target to the exclusion of the set of old targets.

    Well done!!!!! HAHAHA!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!

    You fail again in your misrepresentation. Now you owe me a second apology.

  29. No Sal, read my grade school demonstration:
    More selectively means a bigger difference between best and worst.
    The “exclusion of the set of old targets” has nothing to do with what I wrote in either case.
    When I wrote “sequences that are completely unrelated to any target sequences”, I meant precisely that.
    DNA binding proteins spend a lot for their time bound to completely unrelated sequences, and a decent amount of time unbound, so the amount of time they spend bound to NEAR-matches doesn’t really matter too much.

  30. DNA_Jock:
    No Sal, read my grade school demonstration:
    More selectively means a bigger difference between best and worst.
    The “exclusion of the set of old targets” has nothing to do with what I wrote in either case.
    When I wrote “sequences that are completely unrelated to any target sequences”, I meant precisely that.
    DNA binding proteins spend a lot for their time bound to completely unrelated sequences, and a decent amount of time unbound, so the amount of time they spend bound to NEAR-matches doesn’t really matter too much.

    This is a pedagogical representation of the cis-regulatory region prior to the tandem repeat, with the concsensus binding of the Zinc finger in CAPS:

    1 ACGA aaa

    25 ACGA cga

    ……

    64 ACGA ttt

    after a tandem repeat, will the cis-regulatory regions experience the same amount of binding by the zinc finger protein as before the Zinc Fingers were repeated? Not likely.

    to use DNA_Jock’s words:

    The tandem finger would bind tighter and more selectively

    to #25 more so than the other 63 after the supposed improbable “random tandem” repeat event.

    The problem remains, as I pointed out. About the only criticism now is just mincing of words.

  31. DNA_Jock:
    Oh Sal, I covered this in my cartoon “Grade School” example two days ago.

    LOL!. So #25 will have the same binding affinity as #1, #2,….#64.

    I pointed out:

    KA_25 > KA_1, KA_2…..

    agree or disagree. And if not KA_25, some KA_X. And the point remains.

  32. stcordova,

    You’re grade school example wasn’t what I was talking about. You refuted an argument I didn’t make.

    The fallacy of the Texas sharp shooter fallacy argument second edition 🙂

  33. stcordova: The dimer binding is stronger than the monomer.That was the case in #25 ACGACGA above, or equivalently the dissociation is less for #25.

    For that reason, with the same amount of the transciption factor concentration, more of the transcription will bind to #25 and less to the other 63 cis regulatory regions in my pedagogical example.

    Why would it be the same amount? TFs are themselves subject to regulation, up and down. If the concentration goes down, because more is bound, more can be made.

  34. DNA_Jock is invited to stated for the readers whether believes after the tandem repeat

    KA_25 = KA_1, KA_2, …..KA_64

  35. DNA_Jock is invited to stated for the readers whether believes after the tandem repeat

    KA_25 > KA_1, KA_2, …..KA_64

    and if not KA_25, maybe some KA_X

    Surely DNA_Jock is not asserting after the tandem repeat of the zinc finger protein the relative ranking of the KA_S for each of the cis-regulatory regions will be the same after the tandem repeat?

    And in Jock’s silly grade school example, the teacher should be fired since he asserted Entropy’s silly notion a random tandem will generate 13 zinc fingers covering exactly 84 nucleotides each. Random tandem makes more RANDOM tandems, not orderly tandem repeats.

  36. Here you go entropy, how about you take 1 KRAB domain, slap a solitary zinc finger on it and provide odds that a few iterations of RANDOM ectopic duplication of RANDOM length and RANDOM starting coordinates will make something orderly like:

    http://theskepticalzone.com/wp/wp-content/uploads/2019/04/znf136_zfC2H21.png

    Apparently neither you nor DNA_Jock have even given a guess at the odds.

    I could throw a ZFP with 42 zinc fingers, but that might be really mean. 🙂

  37. Oh Sal, we have covered your “The unreasonable precision of RANDOM TANDEM duplications” previously:

    As I have noted previously, if anyone (other than Sal) does not understand why Entropy is correct about ‘ectopic’ recombination and Sal is hopelessly confused, I will gladly try to explain.
    Here’s a hint: every time Sal uses the word “copy”, he is envisioning a process of copying and pasting. Sal needs to understand cross-over.

    and

    stcordova: Surely DNA_Jock is not asserting after the tandem repeat of the zinc finger protein the relative ranking of the KA_S for each of the cis-regulatory regions will be the same after the tandem repeat?

    Actually, I am asserting that the relative ranking will have changed little, if we are talking about metazoans
    And your “KA_25 is now the best, nyah nyah nyah” “argument” was entirely anticipated by my “look, one of the other sites is now the best binder” exposition in my “grade school” example.
    Please pay attention.

  38. Well, this is awkward.
    I encourage you to read the paper you referenced, Sal, in particular the bit about “search mode”.

    For anyone-who-isn’t-Sal: “Search mode” corresponds to the Egr-1 protein bound to “sequences that are completely unrelated to any target sequences”, like I just explained.
    For Sal, did you understand ANYTHING they wrote about “non-specific complexes”?

  39. So let me describe the fine points as best we understand in the present literature.

    So, what does that mean? Well the zinc finger array will prefer one sequence of DNA over another. It doesn’t mean it won’t bind to a non-specific sequence, but it won’t remain there as long as it will to the target sequence. This is dissociation (KD) and translocation.

    And once the zinc finger array gets to the target sequence, it gives the complex a higher probability of forming and doing it’s business.

    That’s my reading of this:

    http://www.jbc.org/content/288/15/10616.full

    We show here that the DNA binding mode of ZNF217_F67 for the specific and nonspecific DNA sequences is similar and that the observed exchange rate between the free and bound protein is faster for the nonspecific complex, indicative of a higher dissociation rate constant. These observations are consistent with a model in which ZNF217 in the cell probes open genomic DNA with its DNA-binding surface and, by virtue of its slower off-rate when bound to its cognate target sites, is more likely to induce recruitment of additional proteins at such sites. Similar observations have been made for other DNA-binding proteins, including the HoxD9 homeodomain (73, 74) and the ZF protein Zif268 (59). Our study joins a small but growing body of work that begins to provide insight into the mechanism by which proteins can efficiently find their target DNA-binding site immersed in a sea of nonspecific DNA. However, much still remains to be learned about how transcription factors can reach their cognate binding site so quickly in the presence of substantial molecular distractions.

    The point is, whatever random tandem duplication events followed by point mutation happen, the newly evolved protein will preferentially target a different DNA sequence. This preferential targeting can’t happen at random, otherwise it is bad juju.

    One can’t assume, naively as Entropy does, that random tandem ectopic duplications will naturally make zinc finger arrays easily and/or the results will be preferentially selected to go to fixation or evolve further. It is naive as expecting the re-targeting of pre-existing transcription factors to new cis-regulatory sites will necessarily be beneficial. BAD JUJU!

    Or that rare set of random tandem ectopic duplications that are fortuitiously beneficial will emerge frequently enough to will overcome the waiting time problem that several evolutionary biologists (not just a creationist like Dr. Sanford) acknowledge is a real problem.

  40. This point is normally where people would say “and therefore my solution to this problem is that…”

  41. DNA_Jock: sequences that are completely unrelated to any target sequences”, like I just explained.

    Oh I guess you forgot to mention the bind transiently and the TRANSLOCATE to other sequences to actually get to the target sequence in a sea of billions of non-specific sites.

    like I just explained.

    More like obfuscated and distorted. Saving face now, DNA Jock?

    You want to tell the readers with a straight face you expect random tandems to necessarily be functional changes? Do you want to suggest what proportion of such random events will result in functional improvements AND be selectively favorable?

    You want to say with a straight face that binding to non-specific DNA sequences precludes the zinc finger array from eventually parking on the intended target. What the paper I provides shows is that the zinc finger arrays are like people searching for houses to live int. They go into a house briefly and leave if they don’t find what they are looking for. The visit and leave scenario is like zinc finger arrays binding to completely unrelated sequences. But when the target house is found, SHAZAM, the transcription factor finds its home!

  42. OMagain:
    This point is normally where people would say “and therefore my solution to this problem is that…”

    Therefore my solution to this problem is God did it!

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