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 resolution (and the formation) of intermediates in one particular pathway of recombination. The system we are using, the Int-dependent pathway of bacteriophage lambda, is an example of what is called conservative transposition. The proposed experiments depend upon our recent finding that synthetic chi forms of att site DNA, which are formally analogous to Holliday-type recombination intermediates, are resolved by purified Int protein to specific predicted products.
The specific aims of this proposal are: 1) To characterize the resolution of """"""""synthetic"""""""" Holliday intermediates by purified Int protein, with special attention to the questions of specificity and directionality in resolution. 2) To determine how the resolution of Holliday structures is influenced by the accessory proteins IHF and Xis. 3) To determine how the resolution of Holliday structures is influenced by Int protein at sites distant from the region of strand exchange. 4) To study the relationships among the four Int protein binding sites in the Holliday crossover region and the role of DNA:DNA homology in this region. 5) To associate resolution of the Holliday intermediate with a specific domain of Int protein. 6) To find conditions or constructions that favor the isolation of """"""""natural"""""""" Holliday intermediates.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM033928-05
Application #
3284127
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1985-01-01
Project End
1990-03-31
Budget Start
1989-01-01
Budget End
1990-03-31
Support Year
5
Fiscal Year
1989
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