Abstract
Half-lives of Myxococcus xanthus mRNA were determined by inhibiting RNA polymerase with rifampin and then measuring the rate of [35S]methionine incorporation into protein. Vegetative cells resuspended in clone fruiting liquid culture showed an average mRNA half-life of approximately 3.5 min. Developmental cells (24 or 48 hr) exhibited biphasic decay curves with apparent mRNA half-lives of approximately 3.5 min and 20-30 min. The more-stable mRNA species accounted for about 30% of all mRNA present in 24-hr cells and as much as 40% of all mRNA of 48-hr cells. Analysis of the 35S-labeled proteins by NaDodSO4/polyacrylamide gel electrophoresis showed that only 5-10 major polypeptides are synthesized after 30 min of rifampin treatment. One of these is protein S, the spore surface coat protein, because immunoprecipitation of the 35S-labeled proteins with antisera specific for protein S showed continued synthesis of protein S in the presence of rifampin. The half-life for its mRNA was calculated to be 15-30 min. RNA from vegetative and developing cells was pulse labeled with 32PO4 followed by rifampin treatment. Analysis of the labeled RNA on 3% NaDodSO4/polyacrylamide gels showed 5-10 long-lived mRNA bands in developing cells. These results show that there are several abundant mRNA species synthesized developmentally that are exceptionally long lived. The fact that the majority of the mRNA species show the shorter half-life suggests that developing cells retain the normal mechanism for mRNA degradation.
Full text
PDF




Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Campos J. M., Geisselsoder J., Zusman D. R. Isolation of bacteriophage MX4, a generalized transducing phage for Myxococcus xanthus. J Mol Biol. 1978 Feb 25;119(2):167–178. doi: 10.1016/0022-2836(78)90431-x. [DOI] [PubMed] [Google Scholar]
- Chung S., Landfear S. M., Blumberg D. D., Cohen N. S., Lodish H. F. Synthesis and stability of developmentally regulated dictyostelium mRNAs are affected by cell--cell contact and cAMP. Cell. 1981 Jun;24(3):785–797. doi: 10.1016/0092-8674(81)90104-5. [DOI] [PubMed] [Google Scholar]
- GROS F., HIATT H., GILBERT W., KURLAND C. G., RISEBROUGH R. W., WATSON J. D. Unstable ribonucleic acid revealed by pulse labelling of Escherichia coli. Nature. 1961 May 13;190:581–585. doi: 10.1038/190581a0. [DOI] [PubMed] [Google Scholar]
- Gegenheimer P., Watson N., Apirion D. Multiple pathways for primary processing of ribosomal RNA in Escherichia coli. J Biol Chem. 1977 May 10;252(9):3064–3073. [PubMed] [Google Scholar]
- Hagen D. C., Bretscher A. P., Kaiser D. Synergism between morphogenetic mutants of Myxococcus xanthus. Dev Biol. 1978 Jun;64(2):284–296. doi: 10.1016/0012-1606(78)90079-9. [DOI] [PubMed] [Google Scholar]
- Hagen F., Young E. T. Regulation of synthesis of bacteriophage T7 lysozyme mRNA. Virology. 1973 Sep;55(1):231–241. doi: 10.1016/s0042-6822(73)81026-8. [DOI] [PubMed] [Google Scholar]
- Hirashima A., Childs G., Inouye M. Differential inhibitory effects of antibiotics on the biosynthesis of envelope proteins of Escherichia coli. J Mol Biol. 1973 Sep 15;79(2):373–389. doi: 10.1016/0022-2836(73)90012-0. [DOI] [PubMed] [Google Scholar]
- Inouye M., Inouye S., Zusman D. R. Biosynthesis and self-assembly of protein S, a development-specific protein of Myxococcus xanthus. Proc Natl Acad Sci U S A. 1979 Jan;76(1):209–213. doi: 10.1073/pnas.76.1.209. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Inouye M., Inouye S., Zusman D. R. Gene expression during development of Myxococcus xanthus: pattern of protein synthesis. Dev Biol. 1979 Feb;68(2):579–591. doi: 10.1016/0012-1606(79)90228-8. [DOI] [PubMed] [Google Scholar]
- Inouye S., Harada W., Zusman D., Inouye M. Development-specific protein S of Myxococcus xanthus: purification and characterization. J Bacteriol. 1981 Nov;148(2):678–683. doi: 10.1128/jb.148.2.678-683.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ito K., Bassford P. J., Jr, Beckwith J. Protein localization in E. coli: is there a common step in the secretion of periplasmic and outer-membrane proteins? Cell. 1981 Jun;24(3):707–717. doi: 10.1016/0092-8674(81)90097-0. [DOI] [PubMed] [Google Scholar]
- Kaiser D., Manoil C., Dworkin M. Myxobacteria: cell interactions, genetics, and development. Annu Rev Microbiol. 1979;33:595–639. doi: 10.1146/annurev.mi.33.100179.003115. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Levy S. B. Very stable prokaryotic messenger RNA in chromosomeless Escherichia coli minicells. Proc Natl Acad Sci U S A. 1975 Aug;72(8):2900–2904. doi: 10.1073/pnas.72.8.2900. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Linn T., Losick R., Sonenshein A. L. Rifampin resistance mutation of Bacillus subtilis altering the electrophoretic mobility of the beta subunit of ribonucleic acid polymerase. J Bacteriol. 1975 Jun;122(3):1387–1390. doi: 10.1128/jb.122.3.1387-1390.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- NAKADA D., MAGASANIK B. THE ROLES OF INDUCER AND CATABOLITE REPRESSOR IN THE SYNTHESIS OF BETA-GALACTOSIDASE BY ESCHERICHIA COLI. J Mol Biol. 1964 Jan;8:105–127. doi: 10.1016/s0022-2836(64)80153-4. [DOI] [PubMed] [Google Scholar]
- Nelson D. R., Cumsky M. G., Zusman D. R. Localization of myxobacterial hemagglutinin in the periplasmic space and on the cell surface of Myxococcus xanthus during developmental aggregation. J Biol Chem. 1981 Dec 10;256(23):12589–12595. [PubMed] [Google Scholar]
- Rabussay D., Zillig W. A rifampicin resistent rna-polymerase from E. coli altered in the beta-subunit. FEBS Lett. 1969 Oct 21;5(2):104–106. doi: 10.1016/0014-5793(69)80305-4. [DOI] [PubMed] [Google Scholar]
- Rudd K. E., Zusman D. R. RNA polymerase of Myxococcus xanthus: purification and selective transcription in vitro with bacteriophage templates. J Bacteriol. 1982 Jul;151(1):89–105. doi: 10.1128/jb.151.1.89-105.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rudd K., Zusman D. R. Rifampin-resistant mutants of Myxococcus xanthus defective in development. J Bacteriol. 1979 Jan;137(1):295–300. doi: 10.1128/jb.137.1.295-300.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Summers W. C. The process of infection with coliphage T7. IV. Stability of RNA in bacteriophage-infected cells. J Mol Biol. 1970 Aug;51(3):671–678. doi: 10.1016/0022-2836(70)90015-x. [DOI] [PubMed] [Google Scholar]
- Zusman D., Rosenberg E. Division cycle of Myxococcus xanthus. II. Kinetics of stable and unstable ribonucleic acid synthesis. J Bacteriol. 1971 Mar;105(3):801–810. doi: 10.1128/jb.105.3.801-810.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]