Condensation of DNA by spermatid basic nuclear proteins.
Two transition proteins, TP1 and TP2, participate in the repackaging of the spermatid genome early in mammalian spermiogenesis, coincident with the first detectable changes in chromatin condensation. Using an optical trap and a two-channel flow cell to move single DNA molecules into buffer containing protein, we have measured the rates of DNA condensation and decondensation induced by the binding of Syrian hamster transition proteins TP1 and TP2 and protamines P1 and P2. The results show that both transition proteins condense free DNA, with rates similar to those of protamine 1 and 2. DNA molecules condensed with TP1 were significantly less stable than DNA condensed by protamine or by TP2. Experiments conducted with a peptide corresponding to the C-terminal 25 residues of TP2 showed that this domain is responsible for condensing DNA. Experiments conducted with two fragments of TP1 containing arginine and lysine residues demonstrated that DNA binding by TP1 must involve more than these basic sequences. Zinc facilitated the condensation of DNA by P2 but not by TP2. The dissociation rates of TP2 and P2 from DNA were not affected by the addition of zinc.
Brewer
Laurence
L
Electronics Engineering Technologies Division, Lawrence Livermore National Laboratory, Livermore, California 94550, USA. brewer1@llnl.gov
Corzett
Michele
M
Balhorn
Rod
R
D000595:Amino Acid Sequence; D000818:Animals; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000378:metabolism*; D006224:Cricetinae; D004247:DNA / Q000378:metabolism*; D008297:Male; D008647:Mesocricetus; D051379:Mice; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000378:metabolism*; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D011479:Protamines / Q000378:metabolism; D013087:Spermatids / Q000378:metabolism*
D002868:Chromosomal Proteins, Non-Histone; D009687:Nuclear Proteins; D010446:Peptide Fragments; D011479:Protamines; C053886:spermatid transition proteins; D004247:DNA