Processing of a polycistronic mRNA requires a 5′<i>cis</i> element and active translation

Alifano, Pietro; Piscitelli, C; Blasi, Valeria; Rivellini, F.; Nappo, A. G.; Bruni, Carmelo B.; Carlomagno, Maria Stella

doi:10.1111/j.1365-2958.1992.tb01529.x

Cited by 27 publications

(42 citation statements)

References 77 publications

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“…We have reported previously that processing requires an upstream cis-acting element within hisC {Fig. 1A, hatched region) that binds soluble factors in a mobility retardation assay (Alifano et al 1992). This element was able to bind components present in an RNase E-enriched fraction and wild-type but not rnel-derived extracts (data not shown).…”

Section: The Upstream Cis-acting Element Contains Rnase E Target Sitementioning

confidence: 77%

“…This species spans the five distal cistrons of the histidine gene cluster, and its formation requires a cis-acting element located 620 nucleotides upstream of the 5' end of the processed species and the presence of initiating ribosomes at the intercistronic barrier hisC/hisB located immediately downstream (Alifano et al 1992).…”

Section: Mrnamentioning

confidence: 99%

“…This experiment proved the presence of multiple me-dependent cleavage sites within and upstream of the cis-acting element. Deletion of the cis-acting element causes a severe reduction of the 3900-nucleotide processed species [Alifano et al 1992). The evidence provided above suggested a prominent role for this element in binding and subsequent cleavage mediated by RNase E. We tested whether a canonical RNase E target region could substitute for the native element.…”

Section: The Upstream Cis-acting Element Contains Rnase E Target Sitementioning

confidence: 99%

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Ribonuclease E provides substrates for ribonuclease P-dependent processing of a polycistronic mRNA.

Alifano¹,

Rivellini²,

Piscitelli³

et al. 1994

Genes Dev.

144

102

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The polycistronic mRNA of the histidine operon is subject to a processing event that generates a rather stable transcript encompassing the five distal cistrons. The molecular mechanisms by which such a transcript is produced were investigated in Escherichia coil strains carrying mutations in several genes for exo-and endonucleases. The experimental approach made use of S1 nuclease protection assays on in vivo synthesized transcripts, site-directed mutagenesis and construction of chimeric plasmids, dissection of the processing reaction by RNA mobility retardation experiments, and in vitro RNA degradation assays with cellular extracts. We have found that processing requires (1) a functional endonuclease E; (2) target site(s) for this activity in the RNA region upstream of the 5' end of the processed transcript that can be substituted by another well-characterized me-dependent cleavage site; (3) efficient translation initiation of the first cistron immediately downstream of the 5' end; and (4) a functional endonuclease P that seems to act on the processing products generated by ribonuclease E. This is the first evidence that ribonuclease P, an essential ribozyme required for the biosynthesis of tRNA, may also be involved in the segmental stabilization of a mRNA.[Key Words: Escherichia coli; Salmonella typhimurium; his operon; mRNA processing; translation; RNase E; RNase P] Received June 2, 1994; revised version accepted October 18, 1994. mRNA decay in bacteria is mediated by the coordinated action of exonucleases and endonucleases (for review, see Petersen 1992). Polynucleotide phosphorylase (PNPase) and ribonuclease II (RNase II) are the two major 3'--* 5' exonucleases involved in mRNA turnover (Donovan and Kushner 1986). Endoribonuclease III (RNase III) does not affect bulk mRNA stability (Babitzke et al. 1993) and is mostly involved in processing of the 30S rRNA precursor even though RNase III can initiate the functional decay of a few specific mRNAs (Court 1993). Endoribonuclease E (RNase E)was initially characterized as the enzyme that processes the precursor of 5S rRNA (Ghora and Apirion 1978). RNase E is the only endoribonuclease implicated thus far in general mRNA turnover (Arraiano et al. 1988;Mudd et al. 1990b;Babitzke and Kushner 1991). Many mRNAs were shown to contain sites whose cleavage was abolished in strains harboring temperature-sensitive rne mutations at the nonpermissive temperature (Kokoska et al. 1990;Mudd et al. 1990a;Gross 1991 1991; Patel and Dunn 1992;Arraiano et al. 1993;Klug 1993;Mudd and Higgins 1993;Carpousis et al. 1994). rne-dependent cleavages might produce relatively stable processed transcripts, such as the T4 gene 32 (Mudd et al. 1988) and the dicF (Faubladier et al. 1990) and the ghX mRNAs (Brunet al. 1990). In different studies ribonuclease P (RNase P) and other nucleases responsible for stable RNA processing did not seem to be involved in mRNA degradation (Deutscher 1988).We have previously identified in Salmonella typhimurium a rather stable 3900-nucleotide-long mRNA molecule...

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Section: The Upstream Cis-acting Element Contains Rnase E Target Sitementioning

confidence: 77%

Section: Mrnamentioning

confidence: 99%

Section: The Upstream Cis-acting Element Contains Rnase E Target Sitementioning

confidence: 99%

See 1 more Smart Citation

Ribonuclease E provides substrates for ribonuclease P-dependent processing of a polycistronic mRNA.

Alifano¹,

Rivellini²,

Piscitelli³

et al. 1994

Genes Dev.

144

102

View full text Add to dashboard Cite

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“…specific RNase P on the histidine operon mRNA (his mRNA) (1,34), but many examples are known for RNase III and RNase E, and it is interesting to compare them.…”

Section: Distinctions Between Processing and Degradative Rnasesmentioning

confidence: 99%

Processing Endoribonucleases and mRNA Degradation in Bacteria

Kennell

2002

J Bacteriol

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Forty years have passed since the dramatic identification of mRNA, the unstable carrier of genetic information from DNA to protein (15,48,56). During the last decade, there have been scores of papers and reviews that assume that RNase E is the central enzyme for degradation of mRNA. (There have been hundreds of papers and many reviews on RNase E and mRNA degradation, with at least 60 just in the last 2 years. I regret the unintentional omission of worthy ones but only refer to examples in this short presentation.) It is appropriate to consider evidence for and against this conclusion since it bears on our understanding of overall pathways of metabolism. RNase E was identified in 1978 by Apirion et al. (4,44,96) as an endoribonuclease (endo-RNase) that catalyzed the maturation of 5S rRNA by two sequential cleavages at specific sites of the 9S RNA of Escherichia coli. About the same time, Kuwano (recently from training with Apirion) et al. isolated an unusual temperature-sensitive mutant called the ams (for altered mRNA stability) mutant (73,107). About a decade later, it was shown that the ams mutation maps in the gene for RNase E, rne (9,95,99,130). This identification contributed to the now widely held view that RNase E is the principal RNase for initiation of mRNA decay (e.g., see references 33, 34, 35, 49, 57, 87, 122, and 124). FUNCTIONAL DECAY AND MASS DECAY SHOULD BE CONSIDERED SEPARATELYIt is important to recognize the useful compartmentalization of the degradation of an mRNA population for a specific protein (message) into two distinct components: inactivation or loss of function (the ability to synthesize protein) as opposed to loss of mass. It is also important to recognize that decay rates are measured on populations of millions of molecules and are used to infer how a single molecule is degraded. Loss of mass of a heterogeneous population of mRNAs is defined empirically by loss of trichloroacetic acid-precipitable material; the lost fraction generally includes oligonucleotides of less than 12 to 15 nucleotides (nt). For mRNA for a specific protein (message), it has been defined by loss of oligonucleotides that cannot form stable hybrids with their cDNA. The minimal size depends on the stringency of the hybrid reaction and the base composition but is often in the 15-to 30-nt range. A 500-nt mRNA population would only show loss of mass if cleavages occurred near the ends or if it were degraded by an exonuclease. Other terms have been used, such as "bulk decay" and, most commonly, "chemical decay." Mass decay is more specific, since "chemical" could refer to any number of parameters from chemical modifications to structural alterations, as well as loss of mass.Functional decay (inactivation) is defined here by loss of the "capacity" of an mRNA molecule to participate in the initiation of synthesis that leads to the normal functional protein product. It is the rate-limiting event for determining the amount of functional protein from the message, as well as for the onset of rapid degradation of the inact...

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“…The weakness of this promoter allows suppression of the pleiotropic effects of HisH and HisF overexpression by polar mutations upstream p 2 (12), indicating that high levels of HisH and HisF products are not obtained in the absence of constitutive expression from p 1 . However, processed mRNA species derived from the main his transcript are known to have increased stability (1). Thus mutations affecting the production and/or the stability of processed mRNAs containing the distal part of the operon can be expected to affect HisH and HisF levels.…”

mentioning

confidence: 99%

Suppression of the pleiotropic effects of HisH and HisF overproduction identifies four novel loci on the Salmonella typhimurium chromosome: osmH, sfiW, sfiX, and sfiY

Flores

Casadesús

1995

J Bacteriol

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Insertion mutations that suppress some or all the pleiotropic effects of HisH and HisF overproduction were obtained by using transposons Tn10dTet and Tn10dCam. All suppressor mutations proved to be recessive, indicating that their effects were caused by loss of function; thus, the suppressors identify genes that are necessary to trigger the pleiotropic response when HisH and HisF are overproduced. Genetic mapping of the suppressor mutations identifies four novel loci on the Salmonella typhimurium genetic map. Mutations in osmH (min 49) behave as general suppressors that abolish all manifestations of the pleiotropic response. Mutations in sfiY (min 83) suppress cell division inhibition and thermosensitivity but not osmosensitivity. Mutations that suppress only cell division inhibition define another locus, sfiX (min 44). A fourth novel locus, sfiW (min 19), is also involved in cell division inhibition. The phenotype of sfiW mutations is in turn pleiotropic: they suppress cell division inhibition, make S. typhimurium unable to grow in minimal media, and cause slow growth and abnormal colony and cell shape. The inability of sfiW mutants to grow in minimal medium cannot be relieved by any known nutritional requirement or by the use of carbon sources other than glucose. The hierarchy of suppressor phenotypes and the existence of epistatic effects among suppressor mutations suggest a pathwaylike model for the His c pleiotropic response.Constitutive expression of the Salmonella typhimurium histidine operon causes growth inhibition at 42ЊC (24), wrinkled morphology of colonies grown in either 2% glucose or ''green'' plates (28, 42), and growth inhibition on plates containing three to five times the normal concentration of E salts (12, 37). The wrinkled-colony morphology is due to cell filamentation which is also observed in liquid cultures, especially if grown with a high concentration of glucose (25,28,42). This filamentation is sulA independent and unrelated to the SOS response (28).The His c pleiotropic response is caused by increased levels of proteins HisH and HisF (12,25,42). In the close relative Escherichia coli, a similar response is observed when HisH and HisF are overproduced (27). HisH and HisF catalyze the release of 5-amino-4-imidazole carboxamide riboside 5Ј-monophosphate (AICAR), which is also an intermediate in purine biosynthesis (43,44,53). Since AICAR is a potential precursor of the alarmone 5-amino-4-imidazole carboxamide riboside 5Ј-triphosphate (9, 46), several authors had proposed that increased synthesis of AICAR might be responsible for the pleiotropic effects of his overexpression (11,22). However, AICAR does not seem to be involved in the pleiotropic response (25, 26).We have addressed the identification of genes involved in the His c pleiotropic response by the classical approach of studying intergenic (external) suppression (30). Mutations that suppress some or all the manifestations of the pleiotropic response identify four novel loci on the S. typhimurium chromosome: osmH (min 49), sfiW (...

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Processing of a polycistronic mRNA requires a 5′cis element and active translation

Cited by 27 publications

References 77 publications

Ribonuclease E provides substrates for ribonuclease P-dependent processing of a polycistronic mRNA.

Ribonuclease E provides substrates for ribonuclease P-dependent processing of a polycistronic mRNA.

Processing Endoribonucleases and mRNA Degradation in Bacteria

Suppression of the pleiotropic effects of HisH and HisF overproduction identifies four novel loci on the Salmonella typhimurium chromosome: osmH, sfiW, sfiX, and sfiY

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