Requirement for nickel of the transmembrane translocation of NiFe‐hydrogenase 2 in <i>Escherichia coli</i>

Rodrigue, Agnès; Boxer, David H.; Mandrand‐Berthelot, Marie‐Andrée; Wu, Long‐Fei

doi:10.1016/0014-5793(96)00788-0

Cited by 38 publications

(39 citation statements)

References 37 publications

(66 reference statements)

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“…Those early studies identified a strong dependence on FNR for hydrogenase 1 and 2 isoenzyme synthesis (Sawers et al, 1985). Later work suggested that the role of FNR was likely to be indirect and was a reflection of its requirement for nickel transport (Wu et al, 1989 ;Rodrigue et al, 1996). The present study demonstrated only a minor involvement of FNR in transcriptional regulation of hya (see also and hyb operon expression, which is in accord with an indirect involvement of the regulator in operon expression.…”

Section: Discussionsupporting

confidence: 73%

“…However, the results of a later study (Wu et al, 1989) suggested that the reduced level of both enzymes in the fnr mutant was the consequence of impaired nickel transport. Reduced cytoplasmic nickel precludes processing of either hydrogenase precursor and, as a result, the apoenzymes were enzymically inactive and not targeted to the membrane (Rodrigue et al, 1996). An analysis of the transcriptional regulation of the operons encoding both enzymes would resolve this question unequivocally and at the same time provide key information on the control of enzyme synthesis.…”

Section: Introductionmentioning

confidence: 99%

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Transcriptional regulation in response to oxygen and nitrate of the operons encoding the [NiFe] hydrogenases 1 and 2 of Escherichia coli

et al. 1999

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Synthesis of the [NiFe] hydrogenases 1 and 2 of Escherichia coli is induced inresponse to anaerobiosis and is repressed when nitrate is present in the growth medium. The hydrogenase 1 and hydrogenase 2 enzymes are encoded by the polycistronic hyaABCDEF and hybOABCDEFG operons, respectively. Primer extension analysis was used to determine the initiation site of transcription of both operons. This permitted the construction of single-copy lacZ operon fusions, which were used to examine the transcriptional regulation of the two operons. Expression of both was induced by anaerobiosis and repressed by nitrate, which is in complete accord with earlier biochemical studies. Anaerobic induction of the hyb operon was only partially dependent on the FNR protein and, surprisingly, was enhanced by an arcA mutation. This latter result indicated that ArcA suppresses anaerobic hyb expression and that a further factor, which remains to be identified, is involved in controlling anaerobic induction of operon expression. Nitrate repression of hyb expression was mediated by the NarL/NarX and NarP/NarQ two-component regulatory systems. Remarkably, a narP mutant lacked anaerobic induction of hyb expression, even in the absence of added nitrate. Anaerobic induction of hya expression was dependent on the ArcA and AppY regulators, which confirms earlier observations by other authors. Nitrate repression of the hya operon was mediated by both NarL and NarP. Taken together, these data indicate that although the hya and hyb operons share common regulators, there are important differences in the control of expression of the individual operons.

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Section: Discussionsupporting

confidence: 73%

Section: Introductionmentioning

confidence: 99%

Transcriptional regulation in response to oxygen and nitrate of the operons encoding the [NiFe] hydrogenases 1 and 2 of Escherichia coli

et al. 1999

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“…Rocket immunoelectrophoresis was performed as described previously (29). Trypsin-catalyzed solubilization of membrane-bound hydrogenase 2 activity from both washed membranes and spheroplasts was performed essentially as described (30,31). Protein concentration was estimated by the method of Lowry et al (32).…”

Section: Tat-dependent Protein Exportmentioning

confidence: 99%

Sec-independent Protein Translocation in Escherichia coli

Sargent

Stanley

Berks

et al. 1999

Journal of Biological Chemistry

269

120

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In Escherichia coli, transmembrane translocation of proteins can proceed by a number of routes. A subset of periplasmic proteins are exported via the Tat pathway to which proteins are directed by N-terminal "transfer peptides" bearing the consensus (S/T)RRXFLK "twinarginine" motif. The Tat system involves the integral membrane proteins TatA, TatB, TatC, and TatE. Of these, TatA, TatB, and TatE are homologues of the Hcf106 component of the ⌬pH-dependent protein import system of plant thylakoids. Deletion of the tatB gene alone is sufficient to block the export of seven endogenous Tat substrates, including hydrogenase-2. Complementation analysis indicates that while TatA and TatE are functionally interchangeable, the TatB protein is functionally distinct. This conclusion is supported by the observation that Helicobacter pylori tatA will complement an E. coli tatA mutant, but not a tatB mutant. Analysis of Tat component stability in various tat deletion backgrounds shows that TatC is rapidly degraded in the absence of TatB suggesting that TatC complexes, and is stabilized by, TatB.

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“…In addition, certain Tat precursors associate with other polypeptides in the cytoplasm that do not contain signal peptides. In a process called ''hitchhiker export,'' the two polypeptides form a complex that is targeted to the Tat translocation apparatus and then is exported to the periplasm by virtue of the signal peptide in the precursor (Rodrigue et al 1996). In the absence of the assembly partner or when cofactor assembly has been impaired by mutation, protein export through Tat does not take place (Rodrigue et al 1996;Halbig et al 1999).…”

mentioning

confidence: 99%

“…In a process called ''hitchhiker export,'' the two polypeptides form a complex that is targeted to the Tat translocation apparatus and then is exported to the periplasm by virtue of the signal peptide in the precursor (Rodrigue et al 1996). In the absence of the assembly partner or when cofactor assembly has been impaired by mutation, protein export through Tat does not take place (Rodrigue et al 1996;Halbig et al 1999). These observations together with in vitro and genetic evidence point to a protein folding quality control mechanism that is intrinsic to Tat translocation (DeLisa et al 2003).…”

mentioning

confidence: 99%

A bacterial two‐hybrid system based on the twin‐arginine transporter pathway of E. coli

Strauch

Georgiou

2007

Protein Science

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We have developed a bacterial two-hybrid system for the detection of interacting proteins that capitalizes on the folding quality control mechanism of the Twin Arginine Transporter (Tat) pathway. The Tat export pathway is responsible for the membrane translocation of folded proteins, including proteins consisting of more than one polypeptide, only one of which contains a signal peptide (''hitchhiker export''). Here, one protein (bait) is expressed as a fusion to a Tat signal peptide, whereas the second protein (prey) is fused to a protein reporter that can confer a phenotype only after export into the bacterial periplasmic space. Since the prey-reporter fusion lacks a signal peptide, it can only be exported as a complex with the bait-signal peptide fusion that is capable of targeting the Tat translocon. Using maltose-binding protein as a reporter, clones expressing interacting proteins can be grown on maltose minimal media or on MacConkey plates. In addition, we introduce the use of the cysteine disulfide oxidase DsbA as a reporter. Export of a signal peptide-prey:bait-DsbA complex into the periplasm allows complementation of dsbA À mutants and restores the formation of active alkaline phosphatase, which in turn can be detected by a chromogenic assay.Keywords: protein interaction; Tat pathway; protein export; two-hybrid system; folding; fusion protein; maltose-binding protein; DsbA Supplemental material: see www.proteinscience.orgIn recent years, the numbers of genes identified in a broad spectrum of organisms has grown exponentially. A major challenge will be categorizing the corresponding proteins into their functional units within the cell. Several approaches have been used to assign newly identified proteins into cellular networks (Galperin and Koonin 2000; Bork et al. 2004;de Lichtenberg et al. 2005). In vitro, interacting proteins can be detected by mass spectrometry or by chromatographic techniques, typically following genetic fusion with an appropriate affinity tag (Rigaut et al. 1999;Gavin et al. 2002;Butland et al. 2005). In vivo, genetic techniques for the identification of interacting proteins rely mainly on two-hybrid systems and protein complementation assays. A number of such techniques have been developed and used extensively in Escherichia coli (Hu 2001;Karimova et al. 2002Karimova et al. , 2005. So far, the detection of protein interactions in bacteria has capitalized on fusions to transcriptional repressors such as lcI, LexA, or AraC, transcriptional activators (involving the recruitment of RNA polymerase or the dimerization of the Vibrio cholera ToxR), complementation of biosynthetic Reprint requests to: George Georgiou, Chemical Engineering/UT Austin, 1 University Station C0400, Austin, TX 78712-0231, USA; e-mail: gg@che.utexas.edu; fax: (512) 471-7963.Abbreviations: Tat, twin-arginine translocation; Sec, general secretory pathway; TorA, trimethylamine N-oxide reductase; AP, alkaline phosphatase; MBP, maltose-binding protein; Im2, immunity protein 2; E2, endonuclease domain of colici...

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Requirement for nickel of the transmembrane translocation of NiFe‐hydrogenase 2 in Escherichia coli

Cited by 38 publications

References 37 publications

Transcriptional regulation in response to oxygen and nitrate of the operons encoding the [NiFe] hydrogenases 1 and 2 of Escherichia coli

Transcriptional regulation in response to oxygen and nitrate of the operons encoding the [NiFe] hydrogenases 1 and 2 of Escherichia coli

Sec-independent Protein Translocation in Escherichia coli

A bacterial two‐hybrid system based on the twin‐arginine transporter pathway of E. coli

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