Yen Choo

Prof. Yen Choo

orcid
avatar
Co-authors
avatar
Following
avatar
Followers

Research Focus & Metrics

Explore full metrics on Scilit

Academic Career Length

34Years (1993-Present)

(Calculated once Scilit publication data is synced: current year − year of first publication + 1)

Representative Publications

View All Publications
8 November 1994 in Proceedings of the National Academy of Sciences
Verified
Journal Article
Closed access
PDF file
YCYen Choo
Yen Choo
AKA Klug
A Klug
We have used two selection techniques to study sequence-specific DNA recognition by the zinc finger, a small, modular DNA-binding minidomain. We have chosen zinc fingers because they bind as independent modules and so can be linked together in a peptide designed to bind a predetermined DNA site. In this paper, we describe how a library of zinc fingers displayed on the surface of bacteriophage enables selection of fingers capable of binding to given DNA triplets. The amino acid sequences of selected fingers which bind the same triplet are compared to examine how sequence-specific DNA recognition occurs. Our results can be rationalized in terms of coded interactions between zinc fingers and DNA, involving base contacts from a few alpha-helical positions. In the paper following this one, we describe a complementary technique which confirms the identity of amino acids capable of DNA sequence discrimination from these positions.
15 December 1994 in Nature
Verified
Journal Article
Closed access
PDF file
YCYen Choo
Yen Choo
ISIsidro Sanchez-Garcia
Isidro Sanchez-Garcia
AKAaron Klug
Aaron Klug
8 November 1994 in Proceedings of the National Academy of Sciences
Verified
Journal Article
Closed access
PDF file
YCYen Choo
Yen Choo
AKA Klug
A Klug
In the preceding paper [Choo, Y. & Klug, A. (1994) Proc. Natl. Acad. Sci. USA 91, 11163-11167], we showed how selections from a library of zinc fingers displayed on phage yielded fingers able to bind to a number of DNA triplets. Here, we describe a technique to deal efficiently with the converse problem--namely, the selection of a DNA binding site for a given zinc finger. This is done by screening against libraries of DNA triplet binding sites randomized in two positions but having one base fixed in the third position. The technique is applied here to determine the specificity of fingers previously selected by phage display. We find that some of these fingers are able to specify a unique base in each position of the cognate triplet. This is further illustrated by examples of fingers which can discriminate between closely related triplets as measured by their respective equilibrium dissociation constants. Comparing the amino acid sequences of fingers which specify a particular base in a triplet, we infer that in most instances, sequence-specific binding of zinc fingers to DNA can be achieved by using a small set of amino acid-nucleotide base contacts amenable to a code.
1 February 1997 in Current Opinion in Structural Biology
Verified
Journal Article
Closed access
YCYen Choo
Yen Choo
AKAaron Klug
Aaron Klug
1 July 2001 in Nature Biotechnology
Verified
Journal Article
Closed access
PDF file
MIMark Isalan
Mark Isalan
AKAaron Klug
Aaron Klug
YCYen Choo
Yen Choo
27 May 1997 in Proceedings of the National Academy of Sciences
Verified
Journal Article
Open access
PDF file
MIMark Isalan
Mark Isalan
YCYen Choo
Yen Choo
AKAaron Klug
Aaron Klug
Zif268-like zinc fingers are generally regarded as independent DNA-binding modules that each specify three base pairs in adjacent, but discrete, subsites. However, crystallographic evidence suggests that a contact also can occur from the second helical position of one finger to the subsite of the preceding finger. Here we show for the three-finger DNA-binding domain of the protein Zif268, and a panel of variants, that deleting the putative contact from finger 3 can affect the binding specificity for the 5′ base in the adjoining triplet, which forms part of the binding site of finger 2. This finding demonstrates that Zif268-like zinc fingers can specify overlapping 4-bp subsites, and that sequence specificity at the boundary between subsites arises from synergy between adjacent fingers. This has important implications for the design and selection of zinc fingers with novel DNA binding specificities.