Our long-range goal is to achieve a detailed understanding of the molecular mechanisms involved in site-specific recombination. As a model we are studying the Int-dependent recombination pathway of the lambdoid phages, lambda, 080 and P22. The major portion of our effort will continue to focus on the lambda system; a great deal is known about this system and the rate of progress in its analysis has recently been taking some dramatic steps forward. The experiments in this application are intended to complement studies being carried out in other laboratories. For the purpose of presenting our research plan, the aims of this project can be viewed as addressing seven specific questions or problems. The first five are concerned primarily with the general mechanisms of site-specific recombination and the last two concentrate specifically on the remarkable control over directionality in this genetic transaction. 1. What are the limitations on the size of the overlap region? 2. Are the differences (in DNA sequence and affinity for Int) among the four core-type sites an intrinsic feature of the recombination reaction? 3. How do the arm-type Int binding sites function during recombination? 4. What additional insights into the basic mechanisms of site-specific recombination are provided by the features of the 080 and P22 recombination systems? 5. To use gel electrophoresis mobility shifts and electron micro-scopy to study the nature of att site complexes and the kinds of interactions that determine integrative and excisive recombination. 6. What is the minimal set of att site elements necessary for excisive recombination in comparison to the set defined for integrative recombination? 7. What are the roles of the individual arm-type binding sites as (potential) determinants in the control of directionality?

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI013544-10
Application #
3125464
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1977-09-01
Project End
1990-08-31
Budget Start
1986-09-01
Budget End
1987-08-31
Support Year
10
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Brown University
Department
Type
Schools of Medicine
DUNS #
001785542
City
Providence
State
RI
Country
United States
Zip Code
02912
Tirumalai, R S; Kwon, H J; Cardente, E H et al. (1998) Recognition of core-type DNA sites by lambda integrase. J Mol Biol 279:513-27
Nunes-Duby, S E; Kwon, H J; Tirumalai, R S et al. (1998) Similarities and differences among 105 members of the Int family of site-specific recombinases. Nucleic Acids Res 26:391-406
Azaro, M A; Landy, A (1997) The isomeric preference of Holliday junctions influences resolution bias by lambda integrase. EMBO J 16:3744-55
Tirumalai, R S; Healey, E; Landy, A (1997) The catalytic domain of lambda site-specific recombinase. Proc Natl Acad Sci U S A 94:6104-9
Kwon, H J; Tirumalai, R; Landy, A et al. (1997) Flexibility in DNA recombination: structure of the lambda integrase catalytic core. Science 276:126-31
Nunes-Duby, S E; Yu, D; Landy, A (1997) Sensing homology at the strand-swapping step in lambda excisive recombination. J Mol Biol 272:493-508
Tirumalai, R S; Pargellis, C A; Landy, A (1996) Identification and characterization of the N-ethylmaleimide-sensitive site in lambda-integrase. J Biol Chem 271:29599-604
Zahn, K; Landy, A (1996) Modulation of lambda integrase synthesis by rare arginine tRNA. Mol Microbiol 21:69-76
Nunes-Duby, S E; Azaro, M A; Landy, A (1995) Swapping DNA strands and sensing homology without branch migration in lambda site-specific recombination. Curr Biol 5:139-48
Nunes-Duby, S E; Smith-Mungo, L I; Landy, A (1995) Single base-pair precision and structural rigidity in a small IHF-induced DNA loop. J Mol Biol 253:228-42

Showing the most recent 10 out of 34 publications