RNA components of Escherichia coli degradosome: evidence for rRNA decay.

Recently, we found that a multicomponent ribonucleolytic degradosome complex formed around RNase E, a key mRNA-degrading and 9S RNA-processing enzyme, contains RNA in addition to its protein components. Herein we show that the RNA found in the degradosome consists primarily of rRNA fragments that have a range of distinctive sizes. We further show that rRNA degradation is carried out in the degradosome by RNase E cleavage of A+U-rich single-stranded regions of mature 16S and 23S rRNAs. The 5S rRNA, which is known to be generated by RNase E processing of the 9S precursor, was also identified in the degradosome, but tRNAs, which are not cleaved by RNase E in vitro, were absent. Our results, which provide evidence that decay of mature rRNAs occurs in growing Escherichia coli cells in the RNA degradosome, implicate RNase E in degradosome-mediated decay.

[1]  F. Neidhardt The regulation RNA synthesis in bacteria. , 1964, Progress in nucleic acid research and molecular biology.

[2]  D. Schlessinger,et al.  Synthesis and breakdown of ribonucleic acid in Escherichia coli starving for nitrogen. , 1966, Biochimica et biophysica acta.

[3]  A. L. Koch,et al.  Effect of growth rate on the relative rates of synthesis of messenger, ribosomal and transfer RNA in Escherichia coli. , 1972, Journal of molecular biology.

[4]  D. Apirion,et al.  The fate of ribosomes in Escherichia coli cells starved for a carbon source. , 1975, The Journal of biological chemistry.

[5]  K. Gausing Regulation of ribosome production in Escherichia coli: synthesis and stability of ribosomal RNA and of ribosomal protein messenger RNA at different growth rates. , 1977, Journal of molecular biology.

[6]  A. Lassar,et al.  A conditional lethal mutant of Escherichia coli which affects the processing of ribosomal RNA. , 1978, The Journal of biological chemistry.

[7]  D. Apirion,et al.  Structural analysis and in vitro processing to p5 rRNA of a 9S RNA molecule isolated from an rne mutant of E. coli , 1978, Cell.

[8]  D. Apirion Isolation, genetic mapping and some characterization of a mutation in Escherichia coli that affects the processing of ribonuleic acid. , 1978, Genetics.

[9]  S. Y. Lee,et al.  Small Stable RNAs from Escherichia coli: Evidence for the Existence of New Molecules and for a New Ribonucleoprotein Particle Containing 6S RNA , 1978, Journal of bacteriology.

[10]  M. Kuwano,et al.  A conditional lethal mutation in an Escherichia coli strain with a longer chemical lifetime of messenger RNA. , 1979, Journal of molecular biology.

[11]  K. Goldblum,et al.  Inactivation of the ribonucleic acid-processing enzyme ribonuclease E blocks cell division , 1981, Journal of bacteriology.

[12]  T. Tomcsanyi,et al.  Processing enzyme ribonuclease E specifically cleaves RNA I. An inhibitor of primer formation in plasmid DNA synthesis. , 1985, Journal of molecular biology.

[13]  C. Richardson,et al.  A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[14]  B. D. Davis,et al.  Role of ribosome degradation in the death of starved Escherichia coli cells , 1986, Journal of bacteriology.

[15]  S. Cole,et al.  The nucleotide sequence of the malT gene encoding the positive regulator of the Escherichia coli maltose regulon. , 1986, Gene.

[16]  E. Harlow,et al.  Antibodies: A Laboratory Manual , 1988 .

[17]  D. Apirion,et al.  The gene for a small stable RNA (10Sa RNA) of Escherichia coli , 1989, Molecular microbiology.

[18]  H. Sambrook Molecular cloning : a laboratory manual. Cold Spring Harbor, NY , 1989 .

[19]  Stanley N Cohen,et al.  The rate of processing and degradation of antisense RNAI regulates the replication of ColE1-type plasmids in vivo , 1991, Cell.

[20]  R. Srivastava,et al.  Location of the RNA‐processing enzymes RNase III, RNase E and RNase P in the Eschenchia coli cell , 1991, Molecular microbiology.

[21]  H. Inokuchi,et al.  Physical map locations of the genes that encode small stable RNAs in Escherichia coli , 1991, Journal of bacteriology.

[22]  G. Mackie,et al.  Secondary structure of the mRNA for ribosomal protein S20. Implications for cleavage by ribonuclease E. , 1992, The Journal of biological chemistry.

[23]  S. Lin-Chao,et al.  High copy number of the pUC plasmid results from a Rom/Rop‐suppressible point mutation in RNA II , 1992, Molecular microbiology.

[24]  F. Hartl,et al.  Functional interaction of heat shock protein GroEL with an RNase E-like activity in Escherichia coli. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[25]  J. Belasco,et al.  Control of messenger RNA stability. , 1993 .

[26]  C. Higgins,et al.  Escherichia coli endoribonuclease RNase E: autoregulation of expression and site‐specific cleavage of mRNA , 1993, Molecular microbiology.

[27]  R. Gutell,et al.  A compilation of large subunit (23S and 23S-like) ribosomal RNA structures: 1993. , 1992, Nucleic acids research.

[28]  S. Cohen,et al.  The ams-1 and rne-3071 temperature-sensitive mutations in the ams gene are in close proximity to each other and cause substitutions within a domain that resembles a product of the Escherichia coli mre locus , 1993, Journal of bacteriology.

[29]  H. Krisch,et al.  Copurification of E. coli RNAase E and PNPase: Evidence for a specific association between two enzymes important in RNA processing and degradation , 1994, Cell.

[30]  H. Causton,et al.  A protein complex mediating mRNA degradation in Escherichia coli , 1994, Molecular microbiology.

[31]  S. Casaregola,et al.  Cloning and analysis of the entire Escherichia coli ams gene. ams is identical to hmp1 and encodes a 114 kDa protein that migrates as a 180 kDa protein. , 1992, Journal of molecular biology.

[32]  S. Cohen,et al.  Effects of nucleotide sequence on the specificity of rne-dependent and RNase E-mediated cleavages of RNA I encoded by the pBR322 plasmid. , 1994, The Journal of biological chemistry.

[33]  S. Cohen,et al.  A+U content rather than a particular nucleotide order determines the specificity of RNase E cleavage. , 1994, The Journal of biological chemistry.

[34]  G. Björk,et al.  Evidence for an RNA Binding Region in the Escherichia coli processing Endoribonuclease RNase E (*) , 1995, The Journal of Biological Chemistry.

[35]  Stanley N Cohen,et al.  RNA degradation in Escherichia coli regulated by 3' adenylation and 5' phosphorylation , 1995, Nature.

[36]  J. Belasco,et al.  RNase E autoregulates its synthesis by controlling the degradation rate of its own mRNA in Escherichia coli: unusual sensitivity of the rne transcript to RNase E activity. , 1995, Genes & development.

[37]  V. Kaberdin,et al.  Proteins associated with RNase E in a multicomponent ribonucleolytic complex. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[38]  S. Cohen,et al.  The N-terminal domain of the rne gene product has RNase E activity and is non-overlapping with the arginine-rich RNA-binding site. , 1996, Journal of molecular biology.

[39]  V. Kaberdin,et al.  RNase E Cleaves at Multiple Sites in Bubble Regions of RNA I Stem Loops Yielding Products That Dissociate Differentially from the Enzyme* , 1996, The Journal of Biological Chemistry.

[40]  C. Higgins,et al.  A DEAD-box RNA helicase in the Escherichia coli RNA degradosome , 1996, Nature.

[41]  C. Higgins,et al.  Polyphosphate kinase is a component of the Escherichia coli RNA degradosome , 1997, Molecular microbiology.

[42]  K. J. McDowall,et al.  RNase E: still a wonderfully mysterious enzyme , 1997, Molecular microbiology.

[43]  M. Deutscher,et al.  An essential function for the phosphate-dependent exoribonucleases RNase PH and polynucleotide phosphorylase , 1997, Journal of bacteriology.