Abstract
The coding activity of bovine hypothalamic poly A+ mRNA for neurophysin I and II immunoreactive proteins was characterized with respect to size and 5' cap. The mRNA was fractionated by methylmercuric hydroxide agarose gel electrophoresis and subsequently translated in vitro in rabbit reticulocyte lysates. Alternatively, mRNA was fractionated by gel exclusion HPLC and translated in wheat germ extracts. Immunoprecipitated translation products were analyzed by gel exclusion HPLC. Neurophysin-immunospecific protein of approximately 17,000 daltons, the size expected for the neuropeptide hormone-neurophysin precursors, was encoded by mRNA species of two size classes. The smaller class of mRNA's was of the size expected from the size of the precursor proteins. The larger class was 5-10 times larger. The low K+ concentration optimum for translation of unfractionated mRNA encoding neurophysin I immunoreactive proteins and the inability of a cap analogue to inhibit this translation suggest that mRNA species encoding neurophysin I-immunoreactive translation products are incompletely capped. By contrast, the mRNA encoding neurophysin II-immunoreactive products appear to contain a normal cap structure.
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Selected References
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- Bailey J. M., Davidson N. Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Anal Biochem. 1976 Jan;70(1):75–85. doi: 10.1016/s0003-2697(76)80049-8. [DOI] [PubMed] [Google Scholar]
- Brownstein M. J., Russell J. T., Gainer H. Synthesis, transport, and release of posterior pituitary hormones. Science. 1980 Jan 25;207(4429):373–378. doi: 10.1126/science.6153132. [DOI] [PubMed] [Google Scholar]
- Chaiken I. M., Hough C. J. Mapping and isolation of large peptide fragments from bovine neurophysins and biosynthetic neurophysin-containing species by high-performance liquid chromatography. Anal Biochem. 1980 Sep 1;107(1):11–16. doi: 10.1016/0003-2697(80)90484-4. [DOI] [PubMed] [Google Scholar]
- Cordell B., Diamond D., Smith S., Pünter J., Schöne H. H., Goodman H. M. Disproportionate expression of the two nonallelic rat insulin genes in a pancreatic tumor is due to translational control. Cell. 1982 Dec;31(3 Pt 2):531–542. doi: 10.1016/0092-8674(82)90309-9. [DOI] [PubMed] [Google Scholar]
- Fischer E. A., Curd J. G., Chaiken I. M. Preparation of biologically active conjugates of bovine neurophysins and other polypeptides with multi-(poly-D,L-alanyl)--poly-L-lysine and their use to elicit antibodies. Immunochemistry. 1977 Aug;14(8):595–602. doi: 10.1016/0019-2791(77)90155-0. [DOI] [PubMed] [Google Scholar]
- Giudice L. C., Chaiken I. M. Cell-free biosynthesis of different high molecular weight forms of bovine neurophysins I and II coded by hypothalamic mRNA. J Biol Chem. 1979 Dec 10;254(23):11767–11770. [PubMed] [Google Scholar]
- Giudice L. C., Chaiken I. M. Immunological and chemical identification of a neurophysin-containing protein coded by messenger RNA from bovine hypothalamus. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3800–3804. doi: 10.1073/pnas.76.8.3800. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Herson D., Schmidt A., Seal S., Marcus A., van Vloten-Doting L. Competitive mRNA translation in an in vitro system from wheat germ. J Biol Chem. 1979 Sep 10;254(17):8245–8249. [PubMed] [Google Scholar]
- Kozak M. Influence of mRNA secondary structure on binding and migration of 40S ribosomal subunits. Cell. 1980 Jan;19(1):79–90. doi: 10.1016/0092-8674(80)90390-6. [DOI] [PubMed] [Google Scholar]
- Land H., Grez M., Ruppert S., Schmale H., Rehbein M., Richter D., Schütz G. Deduced amino acid sequence from the bovine oxytocin-neurophysin I precursor cDNA. Nature. 1983 Mar 24;302(5906):342–344. doi: 10.1038/302342a0. [DOI] [PubMed] [Google Scholar]
- Land H., Schütz G., Schmale H., Richter D. Nucleotide sequence of cloned cDNA encoding bovine arginine vasopressin-neurophysin II precursor. Nature. 1982 Jan 28;295(5847):299–303. doi: 10.1038/295299a0. [DOI] [PubMed] [Google Scholar]
- Lauber M., Nicolas P., Boussetta H., Fahy C., Béguin P., Camier M., Vaudry H., Cohen P. The Mr 80,000 common forms of neurophysin and vasopressin from bovine neurohypophysis have corticotropin- and beta-endorphin-like sequences and liberate by proteolysis biologically active corticotropin. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6086–6090. doi: 10.1073/pnas.78.10.6086. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Majzoub J. A., Rich A., van Boom J., Habener J. F. Vasopressin and oxytocin mRNA regulation in the rat assessed by hybridization with synthetic oligonucleotides. J Biol Chem. 1983 Dec 10;258(23):14061–14064. [PubMed] [Google Scholar]
- Masse M. J., Desbois-Perichon P., Cohen P. Identification of neurophysin-related proteins in bovine neurosecretory granules. Eur J Biochem. 1982 Oct;127(3):609–617. doi: 10.1111/j.1432-1033.1982.tb06916.x. [DOI] [PubMed] [Google Scholar]
- Nicolas P., Camier M., Lauber M., Masse M. J., Möhring J., Cohen P. Immunological identification of high molecular weight forms common to bovine neurophysin and vasopressin. Proc Natl Acad Sci U S A. 1980 May;77(5):2587–2591. doi: 10.1073/pnas.77.5.2587. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pavlakis G. N., Lockard R. E., Vamvakopoulos N., Rieser L., RajBhandary U. L., Vournakis J. N. Secondary structure of mouse and rabbit alpha- and beta-globin mRNAs: differential accessibility of alpha and beta initiator AUG codons towards nucleases. Cell. 1980 Jan;19(1):91–102. doi: 10.1016/0092-8674(80)90391-8. [DOI] [PubMed] [Google Scholar]
- Payvar F., Schimke R. T. Methylmercury hydroxide enhancement of translation and transcription of ovalbumin and conalbumin mRNA's. J Biol Chem. 1979 Aug 25;254(16):7636–7642. [PubMed] [Google Scholar]
- Rosenior J. C., North W. G., Moore G. J. Putative precursors of vasopressin, oxytocin, and neurophysins in the rat hypothalamus. Endocrinology. 1981 Oct;109(4):1067–1072. doi: 10.1210/endo-109-4-1067. [DOI] [PubMed] [Google Scholar]
- Schmale H., Heinsohn S., Richter D. Structural organization of the rat gene for the arginine vasopressin-neurophysin precursor. EMBO J. 1983;2(5):763–767. doi: 10.1002/j.1460-2075.1983.tb01497.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmale H., Leipold B., Richter D. Cell-free translation of bovine hypothalamic mRNA. Synthesis and processing of the prepro-neurophysin I and II. FEBS Lett. 1979 Dec 15;108(2):311–316. doi: 10.1016/0014-5793(79)80553-0. [DOI] [PubMed] [Google Scholar]
- Sehgal P. B., May L. T., LaForge K. S., Inouye M. Unusually long mRNA species coding for human alpha and beta interferons. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6932–6936. doi: 10.1073/pnas.79.22.6932. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Skup D., Millward S. Reovirus-induced modification of cap-dependent translation in infected L cells. Proc Natl Acad Sci U S A. 1980 Jan;77(1):152–156. doi: 10.1073/pnas.77.1.152. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sonenberg N., Trachsel H., Hecht S., Shatkin A. J. Differential stimulation of capped mRNA translation in vitro by cap binding protein. Nature. 1980 May 29;285(5763):331–333. doi: 10.1038/285331a0. [DOI] [PubMed] [Google Scholar]
- Weber L. A., Hickey E. D., Baglioni C. Influence of potassium salt concentration and temperature on inhibition of mRNA translation by 7-methylguanosine5'-monophosphate. J Biol Chem. 1978 Jan 10;253(1):178–183. [PubMed] [Google Scholar]
- Weber L. A., Hickey E. D., Nuss D. L., Baglioni C. 5'-Terminal 7-methylguanosine and mRNA function: influence of potassium concentration on translation in vitro. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3254–3258. doi: 10.1073/pnas.74.8.3254. [DOI] [PMC free article] [PubMed] [Google Scholar]
