DNA condensation by protamine and arginine-rich peptides: analysis of toroid stability using single DNA molecules.
Both somatic cells and sperm have been shown to take up exogenous DNA, but the frequency of its integration is usually low. Scanning probe microscopy studies of sperm chromatin and synthetic DNA-protamine complexes indicate that the coiling of DNA into toroidal subunits, a process initiated in the maturing spermatid to prepare its genome for delivery into the egg, can be mimicked by simply adding protamine to DNA in vitro. The increased resistance of DNA-protamine complexes to nuclease digestion and their structural similarity to native sperm chromatin suggest that the packaging of DNA by protamine might offer a new approach for improving the efficiency of DNA uptake by sperm. Decondensation experiments performed with individual DNA molecules have provided a direct measure of the stability of toroids produced using salmon protamine and smaller arginine-rich peptides. These experiments show that the arginine content of protamine-related sequences can have a dramatic effect on their rate of dissociation from DNA. This technique and the information it provides can be used to identify protamine analogs that can be bound to DNA to increase the efficiency of its uptake by sperm and other cells.
Balhorn
R
R
Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, Livermore, California 94550, USA.
Brewer
L
L
Corzett
M
M
D001120:Arginine / Q000737:chemistry*; D010582:Bacteriophage lambda / Q000737:chemistry; D002843:Chromatin / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D010455:Peptides / Q000737:chemistry*; D010957:Plasmids; D011479:Protamines / Q000737:chemistry*; D012473:Salmine / Q000737:chemistry
D002843:Chromatin; D010455:Peptides; D011479:Protamines; D004247:DNA; D012473:Salmine; D001120:Arginine