Dynamics of protamine 1 binding to single DNA molecules

被引:58
作者
Brewer, L
Corzett, M
Lau, EY
Balhorn, R
机构
[1] Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94550 USA
[2] Lawrence Livermore Natl Lab, Elect Engn Technol Div, Livermore, CA 94550 USA
关键词
D O I
10.1074/jbc.M303610200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Protamine molecules bind to and condense DNA in the sperm of most vertebrates, packaging the sperm genome in an inactive state until it can be reactivated following fertilization. By using methods that enable the analysis of protamine binding to individual DNA molecules, we have monitored the kinetics of DNA condensation and decondensation by protamine 1 (P1) and synthetic peptides corresponding to specific segments of the bull P1 DNA binding domain. Our results show that the number of clustered arginine residues present in the DNA binding domain is the most important factor affecting the condensation and stability of the DNA-protamine complex prior to the formation of inter-protamine disulfide cross-links. The high affinity of P1 for DNA is achieved by the coordinated binding of three anchoring domains, which together in bull P1 contain 19 Arg residues. The single DNA molecule experiments show that sequences containing two or more anchoring domains have an off-rate that is at least 3 orders of magnitude slower than those containing a single domain. The use of Arg, rather than Lys residues, and the inclusion of Tyr or Phe residues in the hinge regions between anchoring domains provide additional stability to the complex.
引用
收藏
页码:42403 / 42408
页数:6
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