Isolation and characterization of trout testis protamine mRNAs lacking poly (A).
Poly(A)+ protamine mRNA was isolated from trout testis cells in a very pure form, and artificial poly(A)- protamine mRNA molecules were derived from it by enzymatic deadenylation with RNAase H from calf thymus after hybridization with oligo(dT). The deadenylated protamine mRNA was found to be active in a wheat germ cell-free system and yielded a labeled product which co-migrated with authentic protamine. These deadenylated mRNA molecules were subsequently used as markers on denaturing polyacrylamide gels to identify and allow the purification of the poly(A)- protamine components known to exist in vivo in the total cellular poly(A)- RNA. RNA species of molecular weights similar to the enzymatically deadenylated subcomponents of protamine mRNA were observed in the natural poly(A)-RNA population of the testis cells. These naturally occurring poly(A)- protamine mRNAs were isolated by preparative  gel electrophoresis and further characterized by 3H-poly(U) hybridization assay, by hybridization to complementary DNA made against highly purified poly(A)+ protamine mRNA, and by their ability to direct protamine synthesis in a cell-free system.
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D008297:Male; D008970:Molecular Weight; D009693:Nucleic Acid Hybridization; D011061:Poly A; D011479:Protamines / Q000378:metabolism*; D012333:RNA, Messenger / Q000302:isolation & purification* / Q000378:metabolism; D012487:Salmonidae / Q000378:metabolism*; D013737:Testis / Q000032:analysis* / Q000166:cytology; D014337:Trout / Q000378:metabolism*
D011479:Protamines; D012333:RNA, Messenger; D011061:Poly A