Characterization of antibodies against methyl-pppN cap structure: plant U3 small nucleolar RNA is recognized by these antibodies.
In eukaryotes, many small nuclear RNAs contain either a trimethylguanosine cap structure of a gamma-monomethyl (me) cap structure. Previously, we reported the characterization of anti-mepppG antibodies which recognize methyl-capped RNAs with G as the initiation nucleotide. We report here the preparation of antibodies against mepppN cap structure. Anti-mepppN antibodies recognized only mepppN from a mixture of mepppN and pppN and immunoprecipitated mepppA-capped U3 small nucleolar RNA from a mixture of cowpea cell RNAs. These anti-mepppN antibodies recognized methylated nucleoside triphosphates (mepppA, mepppC, mepppG and mepppU) with nearly equal efficiency; however, these antibodies did not recognize methyl phosphate or methylated mononucleotides. These antibodies will be useful in the identification and characterization of all methyl-capped RNAs no matter which is the initiation nucleotide.
Liu
M H
MH
Department of Pharmacology, Baylor College of Medicine, Houston, TX 77030.
Busch
R K
RK
Buckley
B
B
Reddy
R
R
D000974:Antibodies, Antinuclear / Q000276:immunology*; D006367:HeLa Cells; D006801:Humans; D010944:Plants / Q000235:genetics*; D012314:RNA Cap Analogs / Q000276:immunology*; D012342:RNA, Small Nuclear / Q000276:immunology*; D011863:Radioimmunoassay
D000974:Antibodies, Antinuclear; D012314:RNA Cap Analogs; D012342:RNA, Small Nuclear