The Hox genes encode a conserved family of homeodomain-containing transcription factors that specify tissue and cellular identities throughout the animal kingdom. As is typical for homeodomain proteins, they tend to bind a wide range of degenerate, TAAT-containing binding sites, raising the fundamental question of how these factors achieve specificity in vivo. We use a variety of approaches, including whole genome binding studies, in vitro DNA binding assays, X-ray crystallography, in vivo functional tests, and high-throughput DNA specificity measurements, to address this question. Our most recent work (Slattery et al., 2011) reveals that Hox cofactors reveal “latent specificity” that is present in the Hox homeodomain, but cannot be used in the absence of the cofactors. In one specific case (Scr; Joshi et al., 2007) we found that latent specificity allows normally unstructured residues in the Hox protein Scr to read a DNA shape: the width of the minor groove.
Current lab members working on this project: Bill Glassford, Siqian Feng, Judith Kribelbauer, Ryan Loker, and Rebecca Delker (collaborations with Barry Honig, Remo Rohs and Harmen Bussemaker).
Tim Zeiske, Nithya Baburajendran, Anna Kaczynska, Julia Brasch, Arthur G. Palmer III, Lawrence Shapiro, Barry Honig, Richard S. Mann Intrinsic DNA shape accounts for affinity differences between Hox- cofactor binding sites. Cell Reports, Volume 24, Issue 9, 28 August 2018, Pages 2221-2230. August 2018
Chaitanya Rastogi, H. Tomas Rube, Judith F. Kribelbauer, Justin Crocker, Ryan E. Loker, Gabriella D. Martini, Oleg Laptenko, William A. Freed-Pastor, Carol Prives, David L. Stern, Richard S. Mann and Harmen J. Bussemaker Accurate and sensitive quantification of protein-DNA binding affinity. Proceedings of the National Academy of Sciences Apr 2018, 201714376; DOI: 10.1073/pnas.1714376115. April 2018
Judith Kribelbauer, Oleg Laptenko, Siying Chen, Gabriella Martii, William Freed-Pastor, Carol Prives, Richard S. Mann, Harmen Bussemaker Quantitative Analysis of the DNA Methylation Sensitivity of Transcription Factor Complexes Cell Reports. 19, 2383–2395. June 2017
Crocker J, Abe N, Rinaldi L, McGregor AP, Frankel N, Wang S, Alsawadi A, Valenti P, Plaza S, Payre F, Mann RS, Stern DL. Low affinity binding site clusters confer hox specificity and regulatory robustness. Cell. 2015 Jan 15;160(1-2):191-203. doi: 10.1016/j.cell.2014.11.041. Epub 2014 Dec 31. PubMed PMID: 25557079; PubMed Central PMCID: PMC4449256.
Abe N, Dror I, Yang L, Slattery M, Zhou T, Bussemaker HJ, Rohs R, Mann RS. Deconvolving the recognition of DNA shape from sequence. Cell. 2015 Apr 9;161(2):307-18. doi: 10.1016/j.cell.2015.02.008. Epub 2015 Apr 2. PubMed PMID: 25843630; PubMed Central PMCID: PMC4422406.
Merabet S, Mann RS. To Be Specific or Not: The Critical Relationship Between Hox And TALE Proteins. Trends Genet. 2016 Jun;32(6):334-47. doi: 10.1016/j.tig.2016.03.004. Epub 2016 Apr 8. Review. PubMed PMID: 27066866; PubMed Central PMCID: PMC4875764.