Site-directed mutations in the vnd/NK-2 homeodomain. Basis of variations in structure and sequence-specific DNA binding.

The Journal of Biological Chemistry
S WeilerJames A Ferretti

Abstract

Secondary structures, DNA binding properties, and thermal denaturation behavior of six site-directed mutant homeodomains encoded by the vnd/NK-2 gene from Drosophila melanogaster are described. Three single site H52R, Y54M, and T56W mutations, two double site H52R/T56W and Y54M/T56W mutations, and one triple site H52R/Y54M/T56W mutation were investigated. These positions were chosen based on their variability across homeodomains displaying differences in secondary structure and DNA binding specificity. Multidimensional NMR, electrophoretic mobility shift assays, and circular dichroism spectropolarimetry studies were carried out on recombinant 80-amino acid residue proteins containing the homeodomain. Position 56, but more importantly position 56 in combination with position 52, plays an important role in determining the length of the recognition helix. The H52R mutation alone does not affect the length of this helix but does increase the thermal stability. Introduction of site mutations at positions 52 and 56 in vnd/NK-2 does not modify their high affinity binding to the 18-base pair DNA fragment containing the vnd/NK-2 consensus binding sequence, CAAGTG. Site mutations involving position 54 (Y54M, Y54M/T56W, and H52R/Y54M/T56W...Continue Reading

References

Jun 1, 1992·Genes & Development·L Lin, W McGinnis
Dec 3, 1991·Biochemistry·A Laughon
Jun 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·M AffolterW J Gehring
Jul 28, 1989·Biochimica Et Biophysica Acta·M P ScottG W Hartzell
Oct 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·Y Kim, M Nirenberg
Jun 5, 1987·Science·W J Gehring
Dec 1, 1988·Genes & Development·M FraschM Levine
Apr 1, 1995·Protein Engineering·M SuzukiN Yagi
Jul 28, 1995·Journal of Molecular Biology·A DraganescuT D Tullius
Jun 30, 1995·Annals of the New York Academy of Sciences·M NirenbergR Lad
Aug 11, 1995·Journal of Molecular Biology·D H TsaoJ A Ferretti
Aug 11, 1994·Nucleic Acids Research·G DamanteR Di Lauro
Jan 1, 1994·Annual Review of Biochemistry·W J GehringT Bürglin
Jul 29, 1994·Cell·W J GehringK Wüthrich
Sep 15, 1996·Developmental Biology·R P Harvey
Jun 1, 1997·Current Opinion in Genetics & Development·E Boncinelli

❮ Previous
Next ❯

Citations

Feb 24, 2005·Human Genetics·Young-In Chi
Aug 2, 2012·Biochemistry·Lagnajeet PradhanHyun-Joo Nam
Oct 29, 1999·DNA and Cell Biology·M L TejadaR G Deeley
Jun 28, 2000·Nucleic Acids Research·L PellizzariG Damante
Sep 26, 2000·Molecular and Cellular Biology·V DaveJ Ma
Sep 17, 2002·Proceedings of the National Academy of Sciences of the United States of America·Lan-Hsiang WangMarshall Nirenberg
Jun 24, 1998·Proceedings of the National Academy of Sciences of the United States of America·B XiangJ A Ferretti
May 5, 2005·Proceedings of the National Academy of Sciences of the United States of America·Lan-Hsiang WangMarshall Nirenberg
Mar 16, 2007·Proceedings of the National Academy of Sciences of the United States of America·Guoyan ZhaoJames B Skeath
Mar 11, 2003·Proceedings of the National Academy of Sciences of the United States of America·Keita KoizumiJames A Ferretti
Sep 2, 2004·Proceedings of the National Academy of Sciences of the United States of America·Alexander Stepchenko, Marshall Nirenberg
Apr 1, 2015·Proceedings of the National Academy of Sciences of the United States of America·Yohei HayashiRobert J Fletterick
Jan 8, 2008·Journal of Molecular Biology·Ralf JauchPrasanna R Kolatkar
Jan 13, 2006·Journal of Molecular Biology·Jamie M Baird-TitusMark Rance
Nov 15, 2000·Developmental Biology·D B SparrowT J Mohun
Oct 12, 2000·The Journal of Biological Chemistry·R H Harrington, A Sharma
Aug 19, 2003·The Journal of Biological Chemistry·Elliott J StollarBen F Luisi
Jan 4, 2006·Biochemistry·Anatoly I DraganPeter L Privalov

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.