Proteins associated with RNase E in a multicomponent ribonucleolytic complex.
Proceedings of the National Academy of SciencesPublished 30 April 1996Open access
András Miczák, Vladimir R. Kaberdin, Chi Wei, Sue Lin‐Chao
Citations354
Generate an AI Snapshot to get a quick, structured summary of this paper.
Study Snapshot
ObjectiveStudy objective
MethodsResearch methodology
PopulationPopulation studied
Sample sizeSample sizes
OutcomesStudy outcomes here
ResultsStudy results comes here
LimitationsResearch study limitations comes here
A concise AI-generated summary of the paper will appear here once you click Generate AI Snapshot.
TL;DR
N-terminal sequencing of proteins revealed the presence in the RNase E complex not only of polynucleotide phosphorylase but also of DnaK, RNA helicase, and enolase, which indicates the FLAG-Rne complex hasRNase E activity in vivo and in vitro.
Abstract
The Escherichia coli endoribonuclease RNase E is essential for RNA processing and degradation. Earlier work provided evidence that RNase E exists intracellularly as part of a multicomponent complex and that one of the components of this complex is a 3'-to-5' exoribonuclease, polynucleotide phosphorylase (EC 2.7.7.8). To isolate and identify other components of the RNase E complex, FLAG-epitope-tagged RNase E (FLAG-Rne) fusion protein was purified on a monoclonal antibody-conjugated agarose column. The FLAG-Rne fusion protein, eluted by competition with the synthetic FLAG peptide, was found to be associated with other proteins. N-terminal sequencing of these proteins revealed the presence in the RNase E complex not only of polynucleotide phosphorylase but also of DnaK, RNA helicase, and enolase (EC 4.2.1.11). Another protein associated only with epitope-tagged temperature-sensitive (Rne-3071) mutant RNase E but not with the wild-type enzyme is GroEL. The FLAG-Rne complex has RNase E activity in vivo and in vitro. The relative amount of proteins associated with wild-type and Rne-3071 expressed at an elevated temperature differed.
Keywords
Biochemistry, Genetics and Molecular Biology
NatureCleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
251,640 Citations1970Ulrich K. Laemmli
Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products.
Cellular Physiology and BiochemistryShort-Term Effects of Nose-Only Cigarette Smoke Exposure on Glutathione Redox Homeostasis, Cytochrome P450 1A1/2 and Respiratory Enzyme Activities in Mice Tissues
47,283 Citations2013Haider Raza, Annie John +1 more
The results suggest that even short term exposure of cigarette smoke have adverse effects on mitochondrial functions and redox homeostasis in tissues which may progress to further complications associated with chronic smoking.
Proceedings of the National Academy of SciencesA bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes.
3,171 Citations1985Stanley Tabor, C C Richardson
A coupled system that permits the exclusive expression of genes under the control of a T7 RNA polymerase promoter and its use to express high levels of phage T7 gene 5 protein, a subunit of T7 DNA polymerase is described.
Annual Review of BiochemistryMOLECULAR CHAPERONES
1,113 Citations1991R. John Ellis, Saskia M. van der Vies
Proceedings of the National Academy of SciencesMajor heat shock gene of Drosophila and the Escherichia coli heat-inducible dnaK gene are homologous.
516 Citations1984James C.A. Bardwell, Elizabeth A. Craig
Molecular MicrobiologyThe cold‐shock response — a hot topic
439 Citations1994Pamela G. Jones, Masayori Inouye
It has been proposed that the state of the ribosome is the physiological sensor for the induction of cold‐shock proteins and that the Csp proteins, which share sequence similarity with other prokaryotic proteins and with the‘cold‐shock domain’ of eukaryotic Y‐box proteins, may have a function in activating transcription or unwinding or masking RNA molecules.
CellCopurification of E. coli RNAase E and PNPase: Evidence for a specific association between two enzymes important in RNA processing and degradation
430 Citations1994Agamemnon J. Carpousis, Griet Van Houwe +2 more
The discovery of a specific association between RNAase E and PNPase raises the intriguing possibility that these enzymes act cooperatively in the processing and degradation of RNA.
Molecular MicrobiologyRegulation of the <i>Escherichia coli</i> heat‐shock response
340 Citations1993Bernd Bukau
It is proposed that one important signal transduction pathway that converts stress to a heat‐shock response relies on the sequestering of DnaJ through binding to damaged proteins which derepresses and stabilizes σ32.
Annual Review of Biophysics and Biomolecular StructureProtein Folding in the Cell: The Role of Molecular Chaperones Hsp70 and Hsp60
309 Citations1992F. Ulrich Hartl, Jörg Martin +1 more
Multiple Functions of Mitochondrial Hsp60 308 Molecular Mechanism of Chaperon in Action .
CellStructural analysis and in vitro processing to p5 rRNA of a 9S RNA molecule isolated from an rne mutant of E. coli
250 Citations1978Basanta K. Ghora, David Apirion
E. coli contains at least three different sequence variants of 5S rRNA, all of which can be identified in the 9S transcript, indicating that 9S RNA is transcribed from most, if not all, of the active rRNA genes, and that RNAase E processes transcripts derived from all these r RNA genes.
Molecular MicrobiologyA protein complex mediating mRNA degradation in <i>Escherichia coli</i>
223 Citations1994Béatrice Py, Helen C. Causton +2 more
Evidence is presented for a multiprotein complex which includes at least two enzymes that play important roles in mRNA degradation: the exoribonuclease poly‐nucleotide phosphorylase (PNPase) and the endorlbo‐nuclease RNase E. coli.
Molecular MicrobiologyRNase E, an endoribonuclease, has a general role in the chemical decay of <i>Escherichia coli</i> mRNA: evidence that <i>rne</i> and <i>ams</i> are the same genetic locus
217 Citations1990Elisabeth A. Mudd, Henry Krisch +1 more
It is shown that this endoribonuclease has a general role in E. coli mRNA turnover and affects the stability of specific transcripts, and the rne mutation on functional stability of mRNA was much less pronounced than that on chemical stability, although the expression of some genes was affected.
CellThe rate of processing and degradation of antisense RNAI regulates the replication of ColE1-type plasmids in vivo
209 Citations1991Sue Lin‐Chao, Stanley N. Cohen
It is shown that the rate of degradation of RNAI, an anti-sense repressor of the replication primer RNAII, is a key element of control in the replication of ColE1-type plasmids in vivo.
NatureRNA degradation in Escherichia coli regulated by 3' adenylation and 5' phosphorylation
199 Citations1995Fengfeng Xu, Stanley N. Cohen
The discovery that adenylation-mediated degradation by polynucleotide phosphorylase imparts an mRN A-Iike half-life to RNA I suggests a possible mechanism to account for the rapid decay of mRNA10 in E. coli.
The EMBO JournalInvolvement of the chaperonin dnaK in the rapid degradation of a mutant protein in Escherichia coli.
196 Citations1992Sherman MYu, Alfred L. Goldberg
Journal of Molecular BiologyThe N-terminal Domain of the Gene Product has RNase E Activity and is Non-overlapping with the Arginine-rich RNA-binding Site
161 Citations1996Kenneth J. McDowall, Stanley N. Cohen
The results, which map the catalytic domain of RNase E, indicate the existence of discrete functional domains within the multifaceted Rne protein.
Genes & DevelopmentRNase 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.
161 Citations1995Chaitanya Jain, Joel G. Belasco
It is shown that RNase E represses its own synthesis by reducing the cellular concentration of the rne (RNase E) gene transcript, which is a long and highly labile mRNA that can be unusually sensitive to changes in mRNA stability.
NatureSite-specific RNase E cleavage of oligonucleotides and inhibition by stem–loops
144 Citations1995Kenneth J. McDowall, Vladimir R. Kaberdin +3 more
These findings indicate that, contrary to previous proposals, stem–loops do not serve as entry sites for RNase E, but instead limit cleavage at potentially susceptible sites, and that distant conformational changes can also affect enzyme access.
Journal of BacteriologyInactivation of the ribonucleic acid-processing enzyme ribonuclease E blocks cell division
140 Citations1981K Goldblum, D Apririon
These experiments suggest that an early effect of inactivation of the RNA processing enzyme ribonuclease E is a block in cell division.
Journal of Biological ChemistryNucleotide sequence of the pnp gene of Escherichia coli encoding polynucleotide phosphorylase. Homology of the primary structure of the protein with the RNA-binding domain of ribosomal protein S1.
134 Citations1987Philippe Régnier, M. Grunberg‐Manago +1 more
The sequence of a 3030-nucleotide DNA fragment containing the open reading frames coding for ribosomal protein S15 and polynucleotide phosphorylase is presented and the primary structure homologies between a 69-amino-acid stretch of polyn nucleotide phosphORYlase and the four homologous stretches of ribosomic protein S1 which form its RNA binding site are described.
NaturemRNAs can be stabilized by DEAD-box proteins
120 Citations1994Isabelle lost, Marc Dreyfus
It is reported here that when in Escherichia coli the host RNA polymerase is replaced by the eightfold faster bacteriophage T7 enzyme for the transcription of the lacZ gene, the β-galactosidase yield per transcript is depressed 100-fold, but the overexpression of DEAD-box proteins greatly improves this low yield by stabilizing the corresponding transcripts.
Journal of BacteriologyThe 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
119 Citations1993Kenneth J. McDowall, Rachel Hernández +2 more
Computer analysis revealed that the region altered by the mutations has extensive sequence similarity to a predicted gene product from the mre (murein pathway cluster e) locus of E. coli, which has been implicated previously in determining bacterial cell shape.
Journal of Biological ChemistryEffects of nucleotide sequence on the specificity of rne-dependent and RNase E-mediated cleavages of RNA I encoded by the pBR322 plasmid.
118 Citations1994Sue Lin‐Chao, T.T. Wong +2 more
The effects of specific sequence changes introduced by site-directed mutagenesis on the location of ribonucleolytic cleavage near the 5' end of pBR322 RNA I in rne-3071 and congenic rne+ E. coli and on cleavage of RNA I by RNase E in vitro are reported.
Proceedings of the National Academy of SciencesAutoregulation of the Escherichia coli heat shock response by the DnaK and DnaJ heat shock proteins.
113 Citations1993Krzysztof Liberek, Costa Georgopoulos
The formation of an ATP hydrolysis-dependent complex of DnaJ, sigma 32, and DnaK proteins in vitro suggests a possible mechanism for the autoregulation of the heat shock response.
Journal of BacteriologyProperties of <i>Escherichia coli</i> Mutants Deficient in Enzymes of Glycolysis
104 Citations1977Meher Irani, P.K. Maitra
The gluconeogenic requirement of a growing culture of E. coli was shown to be one-twentieth of its total catabolic and anabolic needs, which countered to a large extent the severe repression of beta-galactosidase synthesis that glucose caused in these mutants.
Journal of Molecular BiologyCloning and analysis of the entire Escherichia coli ams gene
101 Citations1992Serge Casarégola, Annick Jacq +6 more
The data indicate that the highly charged C-terminal region of the protein is responsible for its anomalous behaviour when analysed by SDS/PAGE, and is consistent with reports that ams encodes RNaseE, an enzyme required for the processing of several stable RNAs in E. coli.
Molecular Microbiology<i>Escherichia coli</i> endoribonuclease RNase E: autoregulation of expression and site‐specific cleavage of mRNA
98 Citations1993Elisabeth A. Mudd, Christopher F. Higgins
Using antibodies it is shown that the product of the rne gene is a polypeptide of relative mobility 180kDa, which is consistent with RNase K being a proteolytic fragment of RNase E.
The EMBO JournalE.coli polynucleotide phosphorylase expression is autoregulated through an RNase III‐dependent mechanism.
95 Citations1992Murielle Robert-Le Meur, Claude Portier
The results suggest that polynucleotide phosphorylase requires RNase III cleavages to autoregulate the translation of its message and that this 3′ to 5′ processive enzyme could recognize a specific repressor binding site at the 5′ end of pnp mRNA.
Proceedings of the National Academy of SciencesRNase E activity is conferred by a single polypeptide: overexpression, purification, and properties of the ams/rne/hmp1 gene product.
78 Citations1993Robert S. Cormack, Julie Genereaux +1 more
The results demonstrate that RNase E activity is directly attributable to, and is an inherent property of, an RNA-binding protein, the ams/rne/hmp1 gene product, the Ams/Rne/Hmp1 polypeptide.
Journal of Biological ChemistryAnalysis of the altered mRNA stability (ams) gene from Escherichia coli. Nucleotide sequence, transcriptional analysis, and homology of its product to MRP3, a mitochondrial ribosomal protein from Neurospora crassa.
69 Citations1991Félix Claveríe‐Martín, María R. Díaz-Torres +2 more
The product of the altered mRNA stability (ams) gene of Escherichia coli is involved in decay of mRNA, and the complete nucleotide sequence of a 4-kilobase BamHI restriction fragment containing the ams coding sequence was determined.
Molecular MicrobiologyLocation of the RNA‐processing enzymes RNase III, RNase E and RNase P in the <i>Eschenchia coli</i> cell
67 Citations1991András Miczák, Rai Ajit K. Srivastava +1 more
It is shown that even in a normal wild‐type strain of Escherichia coli these enzymes fractionate primarily with the membrane, which suggests that the RNase P protein anchors the RNA catalytic moiety of the enzyme to a larger entity.
Proceedings of the National Academy of SciencesFunctional interaction of heat shock protein GroEL with an RNase E-like activity in Escherichia coli.
61 Citations1993Björn Sohlberg, Urban Lundberg +2 more
Analysis of the relationship between the two nucleases and the heat shock protein revealed that GroEL interacts functionally with anRNase E-like activity but not with an RNase K activity, a groEL mutation affected 9S RNA processing but not ompA mRNA cleavage, RNase E activity could be precipitated with an antibody against GroEL, and a highly purified GroEL preparation contained RNaseE activity butNot RNaseK activity.
PubMedrhlB, a new Escherichia coli K-12 gene with an RNA helicase-like protein sequence motif, one of at least five such possible genes in a prokaryote.
59 Citations1991Miklós Kálmán, Holly S. Murphy +1 more
It is found that rhlB is necessary for viability only in some genetic backgrounds, and the multiplicity of such genes in a prokaryote raises the possibility of important roles for the corresponding class of biologically widespread proteins.
Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular EnzymologyPurification and properties of ribonuclelase E, an RNA-processing enzyme from Escherichia coli
25 Citations1983Monoj K. Roy, David Apirion
It is shown that the cells could contain a factor that enhances RNAase E activity and that the enzyme requires SH groups, free amino groups, and either of the amino acids tryptophan, tyrosine, histidine or methionine for its activity.
BiochimieThe rne gene and ribonuclease E
6 Citations1993András Miczák, David Apirion
The cloned rne+ gene complements temperature sensitive RNase E mutations and directs the synthesis of a polypeptide in vitro and when cells containing the rne plasmid were treated with chloramphenicol, the pre-existingRNase E became less heat labile with time, leading to the suggestion that in the cell RNaseE undergoes post-translational modification(s).