Abstract
I have tested whether selective degradation of mRNA for ribosomal protein S20 of Escherichia coli occurs under conditions for which the expression of S20 is regulated posttranscriptionally. Blot hybridization of total RNA extracted from cultures at different times after addition of rifampin has permitted the estimation of relative levels of the two S20 mRNA species and their half-lives. In a strain harboring a plasmid containing the complete gene for S20, including the transcriptional terminator, moderate posttranscriptional repression of S20 synthesis is accompanied by a substantial increase in the half-lives of both S20 mRNAs relative to those in the haploid parental strain. In an otherwise identical strain in which the transcriptional terminator is deleted from the plasmid-borne S20 genes, the half-life of total S20 mRNA declines more than twofold relative to that in the haploid parent. Thus accelerated decay of the mRNAs for ribosomal protein S20 appears to be an artifact of deletion of the transcriptional terminator, rather than a physiologically significant consequence of translational repression.
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- Belasco J. G., Nilsson G., von Gabain A., Cohen S. N. The stability of E. coli gene transcripts is dependent on determinants localized to specific mRNA segments. Cell. 1986 Jul 18;46(2):245–251. doi: 10.1016/0092-8674(86)90741-5. [DOI] [PubMed] [Google Scholar]
- Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
- Dennis P. P., Nomura M. Regulation of the expression of ribosomal protein genes in Escherichia coli. J Mol Biol. 1975 Sep 5;97(1):61–76. doi: 10.1016/s0022-2836(75)80022-2. [DOI] [PubMed] [Google Scholar]
- Fallon A. M., Jinks C. S., Strycharz G. D., Nomura M. Regulation of ribosomal protein synthesis in Escherichia coli by selective mRNA inactivation. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3411–3415. doi: 10.1073/pnas.76.7.3411. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Geyl D., Böck A. Synthesis of ribosomal proteins in merodiploid strains and in minicells of Escherichia coli. Mol Gen Genet. 1977 Sep 9;154(3):327–334. doi: 10.1007/BF00571290. [DOI] [PubMed] [Google Scholar]
- Gold L., Pribnow D., Schneider T., Shinedling S., Singer B. S., Stormo G. Translational initiation in prokaryotes. Annu Rev Microbiol. 1981;35:365–403. doi: 10.1146/annurev.mi.35.100181.002053. [DOI] [PubMed] [Google Scholar]
- Gorski K., Roch J. M., Prentki P., Krisch H. M. The stability of bacteriophage T4 gene 32 mRNA: a 5' leader sequence that can stabilize mRNA transcripts. Cell. 1985 Dec;43(2 Pt 1):461–469. doi: 10.1016/0092-8674(85)90176-x. [DOI] [PubMed] [Google Scholar]
- Gottesman M., Oppenheim A., Court D. Retroregulation: control of gene expression from sites distal to the gene. Cell. 1982 Jul;29(3):727–728. doi: 10.1016/0092-8674(82)90434-2. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
- Lindahl L., Zengel J. M. Ribosomal genes in Escherichia coli. Annu Rev Genet. 1986;20:297–326. doi: 10.1146/annurev.ge.20.120186.001501. [DOI] [PubMed] [Google Scholar]
- Mackie G. A. Nucleotide sequence of the gene for ribosomal protein S20 and its flanking regions. J Biol Chem. 1981 Aug 10;256(15):8177–8182. [PubMed] [Google Scholar]
- Mackie G. A., Parsons G. D. Tandem promoters in the gene for ribosomal protein S20. J Biol Chem. 1983 Jun 25;258(12):7840–7846. [PubMed] [Google Scholar]
- Mackie G. A. Structure of the DNA distal to the gene for ribosomal protein S20 in Escherichia coli K12: presence of a strong terminator and an IS1 element. Nucleic Acids Res. 1986 Sep 11;14(17):6965–6981. doi: 10.1093/nar/14.17.6965. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mangiarotti G., Schlessinger D. Polyribosome metabolism in Escherichia coli. I. Extraction of polyribosomes and ribosomal subunits from fragile, growing Escherichia coli. J Mol Biol. 1966 Sep;20(1):123–143. doi: 10.1016/0022-2836(66)90122-7. [DOI] [PubMed] [Google Scholar]
- McKenney K., Shimatake H., Court D., Schmeissner U., Brady C., Rosenberg M. A system to study promoter and terminator signals recognized by Escherichia coli RNA polymerase. Gene Amplif Anal. 1981;2:383–415. [PubMed] [Google Scholar]
- Mott J. E., Galloway J. L., Platt T. Maturation of Escherichia coli tryptophan operon mRNA: evidence for 3' exonucleolytic processing after rho-dependent termination. EMBO J. 1985 Jul;4(7):1887–1891. doi: 10.1002/j.1460-2075.1985.tb03865.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nomura M., Gourse R., Baughman G. Regulation of the synthesis of ribosomes and ribosomal components. Annu Rev Biochem. 1984;53:75–117. doi: 10.1146/annurev.bi.53.070184.000451. [DOI] [PubMed] [Google Scholar]
- Olsson M. O., Gausing K. Post-transciptional control of coordinated ribosomal protein synthesis in Escherichia coli. Nature. 1980 Feb 7;283(5747):599–600. doi: 10.1038/283599a0. [DOI] [PubMed] [Google Scholar]
- Panayotatos N., Truong K. Cleavage within an RNase III site can control mRNA stability and protein synthesis in vivo. Nucleic Acids Res. 1985 Apr 11;13(7):2227–2240. doi: 10.1093/nar/13.7.2227. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Parsons G. D., Mackie G. A. Expression of the gene for ribosomal protein S20: effects of gene dosage. J Bacteriol. 1983 Apr;154(1):152–160. doi: 10.1128/jb.154.1.152-160.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmeissner U., McKenney K., Rosenberg M., Court D. Removal of a terminator structure by RNA processing regulates int gene expression. J Mol Biol. 1984 Jun 15;176(1):39–53. doi: 10.1016/0022-2836(84)90381-4. [DOI] [PubMed] [Google Scholar]
- Singer P., Nomura M. Stability of ribosomal protein mRNA and translational feedback regulation in Escherichia coli. Mol Gen Genet. 1985;199(3):543–546. doi: 10.1007/BF00330773. [DOI] [PubMed] [Google Scholar]
- Wirth R., Böck A. Regulation of synthesis of ribosomal protein S20 in vitro. Mol Gen Genet. 1980;178(2):479–481. doi: 10.1007/BF00270504. [DOI] [PubMed] [Google Scholar]
- Wirth R., Littlechild J., Böck A. Ribosomal protein S20 purified under mild conditions almost completely inhibits its own translation. Mol Gen Genet. 1982;188(1):164–166. doi: 10.1007/BF00333013. [DOI] [PubMed] [Google Scholar]
- Zengel J. M., Mueckl D., Lindahl L. Protein L4 of the E. coli ribosome regulates an eleven gene r protein operon. Cell. 1980 Sep;21(2):523–535. doi: 10.1016/0092-8674(80)90490-0. [DOI] [PubMed] [Google Scholar]
- von Gabain A., Belasco J. G., Schottel J. L., Chang A. C., Cohen S. N. Decay of mRNA in Escherichia coli: investigation of the fate of specific segments of transcripts. Proc Natl Acad Sci U S A. 1983 Feb;80(3):653–657. doi: 10.1073/pnas.80.3.653. [DOI] [PMC free article] [PubMed] [Google Scholar]