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. 1989 Nov;86(21):8280–8283. doi: 10.1073/pnas.86.21.8280

Gamma-monomethyl phosphate: a cap structure in spliceosomal U6 small nuclear RNA.

R Singh 1, R Reddy 1
PMCID: PMC298264  PMID: 2813391

Abstract

U6 small nuclear RNA (snRNA), a component of eukaryotic spliceosomes, is required for splicing of nuclear pre-mRNAs. Whereas trimethylguanosine cap-containing U sn-RNAs are transcribed by RNA polymerase II, the U6 RNA is transcribed by RNA polymerase III and contains a nonnucleotide cap structure on its 5' end. We characterized the cap structure of human U6 snRNA and show that the gamma phosphate of the 5' guanosine triphosphate is methylated. The mobilities of in vivo-modified gamma phosphate from the 5' end of HeLa U6 RNA were identical to the synthetic monomethyl phosphate (CH3-O-P) in two-dimensional chromatography and two-dimensional electrophoresis. The cap structure of U6 RNA is distinct from all other cap structures characterized thus far.

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Selected References

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  1. Banerjee A. K. 5'-terminal cap structure in eucaryotic messenger ribonucleic acids. Microbiol Rev. 1980 Jun;44(2):175–205. doi: 10.1128/mr.44.2.175-205.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berget S. M., Robberson B. L. U1, U2, and U4/U6 small nuclear ribonucleoproteins are required for in vitro splicing but not polyadenylation. Cell. 1986 Aug 29;46(5):691–696. doi: 10.1016/0092-8674(86)90344-2. [DOI] [PubMed] [Google Scholar]
  3. Black D. L., Steitz J. A. Pre-mRNA splicing in vitro requires intact U4/U6 small nuclear ribonucleoprotein. Cell. 1986 Aug 29;46(5):697–704. doi: 10.1016/0092-8674(86)90345-4. [DOI] [PubMed] [Google Scholar]
  4. Brow D. A., Guthrie C. Spliceosomal RNA U6 is remarkably conserved from yeast to mammals. Nature. 1988 Jul 21;334(6179):213–218. doi: 10.1038/334213a0. [DOI] [PubMed] [Google Scholar]
  5. Brow D. A., Guthrie C. Splicing a spliceosomal RNA. Nature. 1989 Jan 5;337(6202):14–15. doi: 10.1038/337014a0. [DOI] [PubMed] [Google Scholar]
  6. Brownlee G. G., Sanger F., Barrell B. G. The sequence of 5 s ribosomal ribonucleic acid. J Mol Biol. 1968 Jun 28;34(3):379–412. doi: 10.1016/0022-2836(68)90168-x. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Das G., Henning D., Wright D., Reddy R. Upstream regulatory elements are necessary and sufficient for transcription of a U6 RNA gene by RNA polymerase III. EMBO J. 1988 Feb;7(2):503–512. doi: 10.1002/j.1460-2075.1988.tb02838.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Edery I., Sonenberg N. Cap-dependent RNA splicing in a HeLa nuclear extract. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7590–7594. doi: 10.1073/pnas.82.22.7590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Epstein P., Reddy R., Henning D., Busch H. The nucleotide sequence of nuclear U6 (4.7 S) RNA. J Biol Chem. 1980 Sep 25;255(18):8901–8906. [PubMed] [Google Scholar]
  11. Flanegan J. B., Petterson R. F., Ambros V., Hewlett N. J., Baltimore D. Covalent linkage of a protein to a defined nucleotide sequence at the 5'-terminus of virion and replicative intermediate RNAs of poliovirus. Proc Natl Acad Sci U S A. 1977 Mar;74(3):961–965. doi: 10.1073/pnas.74.3.961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Georgiev O., Mous J., Birnstiel M. L. Processing and nucleo-cytoplasmic transport of histone gene transcripts. Nucleic Acids Res. 1984 Nov 26;12(22):8539–8551. doi: 10.1093/nar/12.22.8539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Grabowski P. J., Sharp P. A. Affinity chromatography of splicing complexes: U2, U5, and U4 + U6 small nuclear ribonucleoprotein particles in the spliceosome. Science. 1986 Sep 19;233(4770):1294–1299. doi: 10.1126/science.3638792. [DOI] [PubMed] [Google Scholar]
  14. Gupta R. C., Randerath E., Randerath K. An improved separation procedure for nucleoside monophosphates on polyethyleneimine-(PEI-)cellulose thin layers. Nucleic Acids Res. 1976 Nov;3(11):2915–2921. doi: 10.1093/nar/3.11.2915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hart R. P., McDevitt M. A., Nevins J. R. Poly(A) site cleavage in a HeLa nuclear extract is dependent on downstream sequences. Cell. 1985 Dec;43(3 Pt 2):677–683. doi: 10.1016/0092-8674(85)90240-5. [DOI] [PubMed] [Google Scholar]
  16. Kiss T., Antal M., Solymosy F. Plant small nuclear RNAs. II. U6 RNA and a 4.5SI-like RNA are present in plant nuclei. Nucleic Acids Res. 1987 Jan 26;15(2):543–560. doi: 10.1093/nar/15.2.543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Konarska M. M., Padgett R. A., Sharp P. A. Recognition of cap structure in splicing in vitro of mRNA precursors. Cell. 1984 Oct;38(3):731–736. doi: 10.1016/0092-8674(84)90268-x. [DOI] [PubMed] [Google Scholar]
  18. Kunkel G. R., Maser R. L., Calvet J. P., Pederson T. U6 small nuclear RNA is transcribed by RNA polymerase III. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8575–8579. doi: 10.1073/pnas.83.22.8575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mattaj I. W. Cap trimethylation of U snRNA is cytoplasmic and dependent on U snRNP protein binding. Cell. 1986 Sep 12;46(6):905–911. doi: 10.1016/0092-8674(86)90072-3. [DOI] [PubMed] [Google Scholar]
  20. Miura K., Tsuda S., Harada F., Ueda T. Chemical modification of cytosine residues of U6 snRNA with hydrogen sulfide (nucleosides and nucleotides. Part 49 [1]). Nucleic Acids Res. 1983 Sep 10;11(17):5893–5901. doi: 10.1093/nar/11.17.5893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mottram J., Perry K. L., Lizardi P. M., Lührmann R., Agabian N., Nelson R. G. Isolation and sequence of four small nuclear U RNA genes of Trypanosoma brucei subsp. brucei: identification of the U2, U4, and U6 RNA analogs. Mol Cell Biol. 1989 Mar;9(3):1212–1223. doi: 10.1128/mcb.9.3.1212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Reddy R., Henning D., Das G., Harless M., Wright D. The capped U6 small nuclear RNA is transcribed by RNA polymerase III. J Biol Chem. 1987 Jan 5;262(1):75–81. [PubMed] [Google Scholar]
  23. Reddy R., Spector D., Henning D., Liu M. H., Busch H. Isolation and partial characterization of dinoflagellate U1-U6 small RNAs homologous to rat U small nuclear RNAs. J Biol Chem. 1983 Nov 25;258(22):13965–13969. [PubMed] [Google Scholar]
  24. Roiha H., Shuster E. O., Brow D. A., Guthrie C. Small nuclear RNAs from budding yeasts: phylogenetic comparisons reveal extensive size variation. Gene. 1989 Oct 15;82(1):137–144. doi: 10.1016/0378-1119(89)90038-3. [DOI] [PubMed] [Google Scholar]
  25. Shatkin A. J. Capping of eucaryotic mRNAs. Cell. 1976 Dec;9(4 Pt 2):645–653. doi: 10.1016/0092-8674(76)90128-8. [DOI] [PubMed] [Google Scholar]
  26. Shatkin A. J. mRNA cap binding proteins: essential factors for initiating translation. Cell. 1985 Feb;40(2):223–224. doi: 10.1016/0092-8674(85)90132-1. [DOI] [PubMed] [Google Scholar]
  27. Silberklang M., Gillum A. M., RajBhandary U. L. Use of in vitro 32P labeling in the sequence analysis of nonradioactive tRNAs. Methods Enzymol. 1979;59:58–109. doi: 10.1016/0076-6879(79)59072-7. [DOI] [PubMed] [Google Scholar]
  28. Skinner H. B., Adams D. S. Nucleotide sequence of Physarum U6 small RNA. Nucleic Acids Res. 1987 Jan 12;15(1):371–371. doi: 10.1093/nar/15.1.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Steitz J. A. "Snurps". Sci Am. 1988 Jun;258(6):56-60, 63. doi: 10.1038/scientificamerican0688-56. [DOI] [PubMed] [Google Scholar]
  30. Tani T., Ohshima Y. The gene for the U6 small nuclear RNA in fission yeast has an intron. Nature. 1989 Jan 5;337(6202):87–90. doi: 10.1038/337087a0. [DOI] [PubMed] [Google Scholar]
  31. Ulmanen I., Broni B., Krug R. M. Influenza virus temperature-sensitive cap (m7GpppNm)-dependent endonuclease. J Virol. 1983 Jan;45(1):27–35. doi: 10.1128/jvi.45.1.27-35.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

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