Abstract

This project is derived from our long term interest in the mechanisms by which genetic material is reassorted and rearranged. We propose to study the formation, properties and resolution of Holliday recombination intermediates. The system we are using, the Int-dependent site-specific pathway of bacteriophage lambda, has been extensively studied at both the genetic and biochemical levels. Because the Holliday intermediate figures prominently in so many recombination pathways, the results obtained in the proposed studies should be of general significance -- in addition to their importance for site-specific recombination, and the (growing) Int family of recombinases. The proposed experiments depend upon our recent finding that this pathway proceeds via a sequential highly ordered pair of reciprocal strand exchanges. They also draw on our demonstration that the appropriate """"""""synthetic"""""""" recombination intermediates afford a very useful tool for studying Holliday junctions. Six specific questions are addressed in this proposal: 1) What is the role of DNA:DNA homology in initiating strand transfer? 2) What are the role and properties of branch migration in resolution? 3) What are the binding patterns and interactions among the Int proteins bound at the Holliday junction? 4) What is the role of arm-type distal sites in the resolution of Holliday junctions? 5) what are the structural features of the Holliday junction in relation to recombination? 6) Does Int-mediated strand transfer have mechanistic features related to illegitimate recombination?

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM033928-06
Application #
3284122
Study Section
Genetics Study Section (GEN)
Project Start
1985-01-01
Project End
1995-03-31
Budget Start
1990-04-01
Budget End
1991-03-31
Support Year
6
Fiscal Year
1990
Total Cost
Indirect Cost

  Institution

Name
Brown University
Department
Type
Schools of Medicine
DUNS #
001785542
City
Providence
State
RI
Country
United States
Zip Code
02912

  Publications

Laxmikanthan, Gurunathan; Xu, Chen; Brilot, Axel F et al. (2016) Structure of a Holliday junction complex reveals mechanisms governing a highly regulated DNA transaction. Elife 5:
Landy, Arthur (2015) The ? Integrase Site-specific Recombination Pathway. Microbiol Spectr 3:MDNA3-0051-2014
Tong, Wenjun; Warren, David; Seah, Nicole E et al. (2014) Mapping the ? Integrase bridges in the nucleoprotein Holliday junction intermediates of viral integrative and excisive recombination. Proc Natl Acad Sci U S A 111:12366-71
Seah, Nicole E; Warren, David; Tong, Wenjun et al. (2014) Nucleoprotein architectures regulating the directionality of viral integration and excision. Proc Natl Acad Sci U S A 111:12372-7
Matovina, Mihaela; Seah, Nicole; Hamilton, Theron et al. (2010) Stoichiometric incorporation of base substitutions at specific sites in supercoiled DNA and supercoiled recombination intermediates. Nucleic Acids Res 38:e175
Warren, David; Laxmikanthan, Gurunathan; Landy, Arthur (2008) A chimeric Cre recombinase with regulated directionality. Proc Natl Acad Sci U S A 105:18278-83
Hazelbaker, Dane; Azaro, Marco A; Landy, Arthur (2008) A biotin interference assay highlights two different asymmetric interaction profiles for lambda integrase arm-type binding sites in integrative versus excisive recombination. J Biol Chem 283:12402-14
Sun, Xingmin; Mierke, Dale F; Biswas, Tapan et al. (2006) Architecture of the 99 bp DNA-six-protein regulatory complex of the lambda att site. Mol Cell 24:569-80
Mumm, Jeffrey P; Landy, Arthur; Gelles, Jeff (2006) Viewing single lambda site-specific recombination events from start to finish. EMBO J 25:4586-95
Radman-Livaja, Marta; Biswas, Tapan; Ellenberger, Tom et al. (2006) DNA arms do the legwork to ensure the directionality of lambda site-specific recombination. Curr Opin Struct Biol 16:42-50

Showing the most recent 10 out of 52 publications