Abstract
The composition of the 5' polyphosphorylated and capped termini of pulse labeled hnRNA from mouse L cells was analyzed by two-dimensional electrophoresis. Purine tri- and diphosphates: pppG, pppA, ppG and ppA; as well as four varieties of cap structure: m7GpppXm, Xm = Gm, (m6) Am, Cm and Um, were detected. With increasing labeling time the relative proportion of hnRNA molecules with tri- and diphosphorylated 5' ends decreases and the relative proportion of capped hnRNA increases, indicating that caps are metabolically more stable than the polyphosphate termini. About half of the hnRNA molecules that are labeled within 2 hr have capped ends. This finding, together with results of earlier kinetic and structural studies, implies that a relatively high proportion of the labeled hnRNA molecules are mRNA precursors. Large hnRNA molecules exhibit a higher proportion of capped ends and a lower proportion of 5'-triphosphate ends as compared to small hnRNA. Given the lower stability of triphosphate termini relative to caps, this result may mean that capping of some hnRNA molecules can occur before the completion of transcription.
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- Bajszár G., Samarina O. P., Georgiev G. P. On the nature of 5' termini in nuclear pre-mRNA of Ehrlich carcinoma cells. Cell. 1976 Oct;9(2):323–332. doi: 10.1016/0092-8674(76)90122-7. [DOI] [PubMed] [Google Scholar]
- Cory S., Adams J. M. The modified 5'-terminal sequences in messenger RNA of mouse myeloma cells. J Mol Biol. 1975 Dec 25;99(4):519–547. doi: 10.1016/s0022-2836(75)80170-7. [DOI] [PubMed] [Google Scholar]
- Davidson E. H., Klein W. H., Britten R. J. Sequence organization in animal DNA and a speculation on hnRNA as a coordinate regulatory transcript. Dev Biol. 1977 Jan;55(1):69–84. doi: 10.1016/0012-1606(77)90320-7. [DOI] [PubMed] [Google Scholar]
- Fedoroff N., Wellauer P. K., Wall R. Intermolecular duplexes in heterogeneous nuclear RNA from HeLa cells. Cell. 1977 Apr;10(4):597–610. doi: 10.1016/0092-8674(77)90092-7. [DOI] [PubMed] [Google Scholar]
- Furuichi Y., Shatkin A. J. Differential synthesis of blocked and unblocked 5'-termini in reovirus mRNA: effect of pyrophosphate and pyrophosphatase. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3448–3452. doi: 10.1073/pnas.73.10.3448. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hames B. D., Perry R. P. Homology relationship between the messenger RNA and heterogeneous nuclear RNA of mouse L cells. A DNA excess hybridization study. J Mol Biol. 1977 Jan 25;109(3):437–453. doi: 10.1016/s0022-2836(77)80022-3. [DOI] [PubMed] [Google Scholar]
- Perry R. P., Bard E., Hames B. D., Kelly D. E., Schibler U. The relationship between hnRNA and mRNA. Prog Nucleic Acid Res Mol Biol. 1976;19:275–292. doi: 10.1016/s0079-6603(08)60925-3. [DOI] [PubMed] [Google Scholar]
- Perry R. P., Kelley D. E. Kinetics of formation of 5' terminal caps in mRNA. Cell. 1976 Jul;8(3):433–442. doi: 10.1016/0092-8674(76)90156-2. [DOI] [PubMed] [Google Scholar]
- Perry R. P., Kelley D. E., LaTorre J. Synthesis and turnover of nuclear and cytoplasmic polyadenylic acid in mouse L cells. J Mol Biol. 1974 Jan 25;82(3):315–331. doi: 10.1016/0022-2836(74)90593-2. [DOI] [PubMed] [Google Scholar]
- Salditt-Georgieff M., Jelinek W., Darnell J. E., Furuichi Y., Morgan M., Shatkin A. Methyl labeling of HeLa cell hnRNA: a comparison with mRNA. Cell. 1976 Feb;7(2):227–237. doi: 10.1016/0092-8674(76)90022-2. [DOI] [PubMed] [Google Scholar]
- Schibler U., Perry R. P. Characterization of the 5' termini of hn RNA in mouse L cells: implications for processing and cap formation. Cell. 1976 Sep;9(1):121–130. doi: 10.1016/0092-8674(76)90058-1. [DOI] [PubMed] [Google Scholar]
- Schmincke C. D., Herrmann K., Hausen P. Size of primary transcripts in Ehrlich ascites cells as measured by tetraphosphate determination. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1994–1998. doi: 10.1073/pnas.73.6.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Volckaert G., Jou W. M., Fiers W. Analysis of 32P-labeled bacteriophage MS2 RNA by a mini-fingerprinting procedure. Anal Biochem. 1976 May 7;72:433–446. doi: 10.1016/0003-2697(76)90551-0. [DOI] [PubMed] [Google Scholar]
- Wei C., Moss B. 5'-Terminal capping of RNA by guanylyltransferase from HeLa cell nuclei. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3758–3761. doi: 10.1073/pnas.74.9.3758. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Winicov I., Perry R. P. Synthesis methylation, and capping of nuclear RNA by a subcellular system. Biochemistry. 1976 Nov 16;15(23):5039–5046. doi: 10.1021/bi00668a014. [DOI] [PubMed] [Google Scholar]