Formation of native-like mammalian sperm cell chromatin with folded bull protamine.
The DNA of most vertebrate sperm cells is packaged by protamines. The primary structure of mammalian protamine I can be divided into three domains, a central DNA binding domain that is arginine-rich and amino- and carboxyl-terminal domains that are rich in cysteine residues. In native bull sperm chromatin, intramolecular disulfide bonds hold the terminal domains of bull protamine folded back onto the central DNA binding domain, whereas intermolecular disulfide bonds between DNA-bound protamines help stabilize the chromatin of mature mammalian sperm cells. Folded bull protamine was used to condense DNA in vitro under various solution conditions. Using transmission electron microscopy and light scattering, we show that bull protamine forms particles with DNA that are morphologically similar to the subunits of native bull sperm chromatin. In addition, the stability provided by intermolecular disulfide bonds formed between bull protamine molecules within in vitro DNA condensates is comparable with that observed for native bull sperm chromatin. The importance of the bull protamine terminal domains in controlling the bull sperm chromatin morphology is indicated by our observation that DNA condensates formed under identical conditions with a fish protamine, which lacks cysteine-rich terminal domains, do not produce as uniform structures as bull protamine. A model is also presented for the bull protamine.DNA complex in native sperm cell chromatin that provides an explanation for the positions of the cysteine residues in bull protamine that form intermolecular disulfide bonds.
School of Chemistry and Biochemistry, Parker H. Petit Institute of Bioengineering and Biosciences, Georgia Institute of Technology, Atlanta, GA 30332-0400, USA.
Igor D
ID
Vilfan
Christine C
CC
Conwell
Nicholas V
NV
Hud
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000737:chemistry; D002417:Cattle; D002843:Chromatin / Q000737:chemistry / Q000378:metabolism; D003545:Cysteine / Q000737:chemistry; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D007477:Ions; D008297:Male; D008623:Mercaptoethanol / Q000494:pharmacology; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D011479:Protamines / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D012474:Salmon; D013094:Spermatozoa / Q000378:metabolism*; D013997:Time Factors
D002843:Chromatin; D004220:Disulfides; D007477:Ions; D011479:Protamines; D008623:Mercaptoethanol; D004247:DNA; D001120:Arginine; D003545:Cysteine