Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1992 Aug 25;20(16):4299–4304. doi: 10.1093/nar/20.16.4299

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

M H Liu 1, R K Busch 1, B Buckley 1, R Reddy 1
PMCID: PMC334139  PMID: 1508721

Abstract

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.

Full text

PDF
4299

Images in this article

4300Image
on p.4300
4300Image
on p.4300
4301Image
on p.4301
4302Image
on p.4302
4303Image
on p.4303
4303Image
on p.4303

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abel S., Kiss T., Solymosy F. Molecular analysis of eight U1 RNA gene candidates from tomato that could potentially be transcribed into U1 RNA sequence variants differing from each other in similar regions of secondary structure. Nucleic Acids Res. 1989 Aug 11;17(15):6319–6337. doi: 10.1093/nar/17.15.6319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bringmann P., Lührmann R. Purification of the individual snRNPs U1, U2, U5 and U4/U6 from HeLa cells and characterization of their protein constituents. EMBO J. 1986 Dec 20;5(13):3509–3516. doi: 10.1002/j.1460-2075.1986.tb04676.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Busch H., Reddy R., Rothblum L., Choi Y. C. SnRNAs, SnRNPs, and RNA processing. Annu Rev Biochem. 1982;51:617–654. doi: 10.1146/annurev.bi.51.070182.003153. [DOI] [PubMed] [Google Scholar]
  4. Gupta S., Busch R. K., Singh R., Reddy R. Characterization of U6 small nuclear RNA cap-specific antibodies. Identification of gamma-monomethyl-GTP cap structure in 7SK and several other human small RNAs. J Biol Chem. 1990 Nov 5;265(31):19137–19142. [PubMed] [Google Scholar]
  5. Inouye H., Fuchs S., Sela M., Littauer U. Z. Anti-inosine antibodies. Biochim Biophys Acta. 1971 Jul 29;240(4):594–603. doi: 10.1016/0005-2787(71)90717-9. [DOI] [PubMed] [Google Scholar]
  6. Krol A., Ebel J. P., Rinke J., Luhrmann R. U1, U2 and U5 small nuclear RNAs are found in plants cells. Complete nucleotide sequence of the U5 RNA family from pea nuclei. Nucleic Acids Res. 1983 Dec 20;11(24):8583–8594. doi: 10.1093/nar/11.24.8583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Lerner M. R., Steitz J. A. Antibodies to small nuclear RNAs complexed with proteins are produced by patients with systemic lupus erythematosus. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5495–5499. doi: 10.1073/pnas.76.11.5495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Luhrmann R., Appel B., Bringmann P., Rinke J., Reuter R., Rothe S., Bald R. Isolation and characterization of rabbit anti-m3 2,2,7G antibodies. Nucleic Acids Res. 1982 Nov 25;10(22):7103–7113. doi: 10.1093/nar/10.22.7103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Montzka K. A., Steitz J. A. Additional low-abundance human small nuclear ribonucleoproteins: U11, U12, etc. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8885–8889. doi: 10.1073/pnas.85.23.8885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Reddy R., Li W. Y., Henning D., Choi Y. C., Nohga K., Busch H. Characterization and subcellular localization of 7-8 S RNAs of Novikoff hepatoma. J Biol Chem. 1981 Aug 25;256(16):8452–8457. [PubMed] [Google Scholar]
  11. Shumyatsky G. P., Tillib S. V., Kramerov D. A. B2 RNA and 7SK RNA, RNA polymerase III transcripts, have a cap-like structure at their 5' end. Nucleic Acids Res. 1990 Nov 11;18(21):6347–6351. doi: 10.1093/nar/18.21.6347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Singh R., Reddy R. Gamma-monomethyl phosphate: a cap structure in spliceosomal U6 small nuclear RNA. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8280–8283. doi: 10.1073/pnas.86.21.8280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wassarman D. A., Steitz J. A. Structural analyses of the 7SK ribonucleoprotein (RNP), the most abundant human small RNP of unknown function. Mol Cell Biol. 1991 Jul;11(7):3432–3445. doi: 10.1128/mcb.11.7.3432. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES