Abstract
A highly charged component can be isolated from a total RNase T2 digest of nuclear polyadenylylated RNA from HeLa cells that is separable from caps by (dihydroxyboryl)aminoethyl-cellulose chromatography. Chemical and enzymatic analyses show that the component contains a 2′-5′ phosphodiester bond that creates a branch at the 2′-hydroxyl group of one nucleotide already linked to an adjoining nucleotide through the usual 3′-5′ phosphodiester bond. [Formula: see text] This structure was confirmed by analysis of a similar component isolated from nuclease P1 digests of the same nuclear polyadenylylated RNA. Branches occur in roughly 10% of nuclear polyadenylylated RNAs, including those >10S in size, but are absent from cytoplasmic polyadenylylated RNA. Possible implications for branches as intermediates in mRNA processing are discussed.
Keywords: heterogeneous nuclear RNA, mRNA, HeLa cell, RNA processing
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Brawerman G. The Role of the poly(A) sequence in mammalian messenger RNA. CRC Crit Rev Biochem. 1981;10(1):1–38. doi: 10.3109/10409238109114634. [DOI] [PubMed] [Google Scholar]
- Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
- Darnell J. E., Jr Variety in the level of gene control in eukaryotic cells. Nature. 1982 Jun 3;297(5865):365–371. doi: 10.1038/297365a0. [DOI] [PubMed] [Google Scholar]
- Konarska M., Filipowicz W., Domdey H., Gross H. J. Formation of a 2'-phosphomonoester, 3',5'-phosphodiester linkage by a novel RNA ligase in wheat germ. Nature. 1981 Sep 10;293(5828):112–116. doi: 10.1038/293112a0. [DOI] [PubMed] [Google Scholar]
- Konarska M., Filipowicz W., Gross H. J. RNA ligation via 2'-phosphomonoester, 3'5'-phosphodiester linkage: requirement of 2',3'-cyclic phosphate termini and involvement of a 5'-hydroxyl polynucleotide kinase. Proc Natl Acad Sci U S A. 1982 Mar;79(5):1474–1478. doi: 10.1073/pnas.79.5.1474. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Korwek E. L., Nakazato N., Edmonds M., Venkatesan S. Poly(uridylic acid) sequences in messenger ribonucleic acid of HeLa cells. Biochemistry. 1976 Oct 19;15(21):4643–4649. doi: 10.1021/bi00666a015. [DOI] [PubMed] [Google Scholar]
- Nakazato H., Edmonds M. Purification of messenger RNA and heterogeneous nuclear RNA containing poly(a) sequences. Methods Enzymol. 1974;29:431–443. doi: 10.1016/0076-6879(74)29035-9. [DOI] [PubMed] [Google Scholar]
- Penman S. RNA metabolism in the HeLa cell nucleus. J Mol Biol. 1966 May;17(1):117–130. doi: 10.1016/s0022-2836(66)80098-0. [DOI] [PubMed] [Google Scholar]
- SEBRING E. D., SALZMAN N. P. AN IMPROVED PROCEDURE FOR MEASURING THE DISTRIBUTION OF P32O4--AMONG THE NUCLEOTIDES OF RIBONUCLEIC ACID. Anal Biochem. 1964 May;8:126–129. doi: 10.1016/0003-2697(64)90177-0. [DOI] [PubMed] [Google Scholar]
- Sninsky J. J., Last J. A., Gilham P. T. The use of terminal blocking groups for the specific joining of oligonucleotides in RNA ligase reactions containing equimolar concentrations of acceptor and donor molecules. Nucleic Acids Res. 1976 Nov;3(11):3157–3166. doi: 10.1093/nar/3.11.3157. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Venkatesan S., Nakazato H., Kopp D. W., Edmonds M. Properties of a small transcribed poly A sequence in heterogeneous nuclear RNA of HeLa cells. Nucleic Acids Res. 1979 Mar;6(3):1097–1110. doi: 10.1093/nar/6.3.1097. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wallace J. C., Wood W. M., Edmonds M. 5'-Terminal cap structures of oligo(uridylic acid)-containing messenger ribonucleic acid from HeLa cells: comparison with other ribonucleic acid subpopulations. Biochemistry. 1981 Sep 15;20(19):5364–5368. doi: 10.1021/bi00522a002. [DOI] [PubMed] [Google Scholar]