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Nucleic Acids Research, 1992, Vol. 20, No. 16 4299-4304
© 1992

MOLECULAR BIOLOGY

Characterization of antibodies against methyl-pppN cap structure: plant U3 small nucleolar RNA is recognized by these antibodies

Mei-Hua Liu, Rose K. Busch, Becky Buckley1 and Ram Reddy*

Department of Pharmacology Houston, TX 77030, USA 1Division of Molecular Virology, Baylor College of Medicine Houston, TX 77030, USA

*To whom correspondence should be addressed

Received April 16, 1992. Revised July 24, 1992. Accepted July 24, 1992.

In eukaryotes, many small nuclear RNAs contain either a trimethylguanosine cap structure or a {gamma}-monomethyl (me) cap structure. Previously, we reported the characterization of anti-mepppG antibodies which ecognize 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 Immunopreclpltated mepppAcapped U3 small nucleolar RNA from a mixture of cowpea cell RNAs. These anti-mepppN antibodies recognized methylated nucleoslde 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.


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