title: 5'-Terminal and internal methylated nucleosides in herpes simplex virus type 1 mRNA.
abstract: RNA labeled with [methyl-3H]methionine and/or [32P]orthophosphate was isolated from the polyribosomes of herpes simplex virus (HSV) types 1-infected cells and separated into polyadenylylated [poly(A+)]and non-polyadenylylated [poly(A-)] fractions. Virus-specific RNA was obtained by hybridization in liquid to either excess HSV DNA or filters containing immobilized HSV DNA. Analysis in denaturing sucrose gradients indicated that HSV-specific poly(A+) RNA sedimented in a broad peak, with a modal S value of 20. The ratio of [3H]methyl to 32P decreased with increasing size of RNA, suggesting that each RNA chain contains a similar sumber of methyl groups. Further analysis indicated an average of one RNase-resistant structure of the type m7G(5')pppNmpNp or m7G(5')pppNmpNmpNp per 2,780 nucleotides. The following components were identified in the 5'-terminal oligonucleotides of polyribosome-associated HSV-specific poly(A+) and poly(A-) RNA: 7-methylguanosine, N6,2'-O-dimethyladenosine, and the 2'-O-methyl derivatives of guanosine, adenosine, uridine, and denosine, and the 2'-O-methyl derivatives of guanosine, adenosine, uridine, and cytidine. The most common 5'-terminal sequences were m7G(5')pppm6Am and m7G(5')pppGm. An additional modified nucleoside, N6-methyladenosine, was present in an internal position of HSV-specific RNA.
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives / Q000032:analysis; D001483:Base Sequence; D003562:Cytidine / Q000031:analogs & derivatives / Q000032:analysis; D006151:Guanosine / Q000031:analogs & derivatives / Q000032:analysis; D006367:HeLa Cells; D008745:Methylation; D009705:Nucleosides / Q000032:analysis*; D011061:Poly A / Q000032:analysis; D011132:Polyribosomes / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis*; D012367:RNA, Viral / Q000032:analysis*; D018139:Simplexvirus / Q000032:analysis*; D014529:Uridine / Q000031:analogs & derivatives / Q000032:analysis
chemical_list: D009705:Nucleosides; D012333:RNA, Messenger; D012367:RNA, Viral; D006151:Guanosine; D011061:Poly A; D003562:Cytidine; D000241:Adenosine; D014529:Uridine