A Klebsiella aerogenes moaEF operon is controlled by the positive MoaR regulator of the monoamine regulon

Azakami, Hiroyuki; Sugino, Hiroyuki; Iwata, Nobuhide; Yokoro, Natsuko; Yamashita, Mitsuo; Murooka, Yoshikatsu

doi:10.1016/0378-1119(95)00400-z

Cited by 8 publications

(5 citation statements)

References 13 publications

(6 reference statements)

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“…Extrapolated to 100% FGly content, activities of 152 or 205 units/mg, respectively, are calculated for the two protein preparations. These results rule out that AtsB acts as a transcriptional activator, as had been suggested originally (19,20). AtsB rather plays an essential role in the posttranslational oxidation of a conserved serine to FGly.…”

Section: Discussioncontrasting

confidence: 39%

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The Iron Sulfur Protein AtsB Is Required for Posttranslational Formation of Formylglycine in the Klebsiella Sulfatase

Szameit

Miech

Balleininger

et al. 1999

Journal of Biological Chemistry

119

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The catalytic residue of eukaryotic and prokaryotic sulfatases is a ␣-formylglycine. In the sulfatase of Klebsiella pneumoniae the formylglycine is generated by posttranslational oxidation of serine 72. We cloned the atsBA operon of K. pneumoniae and found that the sulfatase was expressed in inactive form in Escherichia coli transformed with the structural gene (atsA). Coexpression of the atsB gene, however, led to production of high sulfatase activity, indicating that the atsB gene product plays a posttranslational role that is essential for the sulfatase to gain its catalytic activity. This was verified after purification of the sulfatase from the periplasm of the cells. Peptide analysis of the protein expressed in the presence of AtsB revealed that half of the polypeptides carried the formylglycine at position 72, while the remaining polypeptides carried the encoded serine. The inactive sulfatase expressed in the absence of AtsB carried exclusively serine 72, demonstrating that the atsB gene is required for formylglycine modification. This gene encodes a 395-amino acid residue iron sulfur protein that has a cytosolic localization and is supposed to directly or indirectly catalyze the oxidation of the serine to formylglycine.

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Section: Discussioncontrasting

confidence: 39%

“…In the present study we characterized this regulation in more detail. Data presented here show that regulation by AtsB is not due to a function as a transcriptional activator, as had been suggested originally (19,20). AtsB rather plays a crucial role in a posttranslational event, namely the conversion of serine 72 to FGly.…”

mentioning

confidence: 56%

The Iron Sulfur Protein AtsB Is Required for Posttranslational Formation of Formylglycine in the Klebsiella Sulfatase

Szameit

Miech

Balleininger

et al. 1999

Journal of Biological Chemistry

119

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“…The region of the M. smegmatis chromosome containing the lon gene is schematized in Figure 1. Sequences upstream of lon encode a homolog from the common mycobacterial antigen family, which encode mycoyl-transferases (36) [most closely resembling antigen 85-C (37)] and a gene likely to encode a dehydrogenase [most closely resembling the ribitol dehydrogenase of Klebsiella aerogenes (38)]. Sequences downstream of lon neither encode significant open reading frames nor potential stemloop structures, characteristic of factor-independent transcriptional terminators.…”

Section: Resultsmentioning

confidence: 99%

The Lon Protease from Mycobacterium smegmatis: Molecular Cloning, Sequence Analysis, Functional Expression, and Enzymatic Characterization

et al. 1998

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We have charterized a Mycobacterium smegmatis gene encoding a homolog of the ATP-dependent protease Lon (La). Our identification of a Lon homolog, in conjunction with our previous work, identifies M. smegmatis as the first known example of a eubacterium containing both Lon and a complete 20S proteasome (containing both alpha- and beta-subunits). Despite the significant primary sequence divergence between M. smegmatis Lon (Ms-Lon) and E. coli Lon (Ec-Lon), expression of Ms-Lon was only moderately toxic to E. coli cells. The ability of E. coli cells to tolerate expression of Ms-Lon reveals that Ms-Lon does not recognize and degrade essential E. coli proteins. We conclude that discrimination against nonsubstrate proteins is broadly conserved between Ec-Lon and Ms-Lon. Additional conservation of substrate recognition was demonstrated by the ability of Ms-Lon to degrade efficiently RcsA, a natural substrate of Ec-Lon. Purified Ms-Lon displays chymotrypsin-like specificity in peptidase assays that are stimulated by unfolded protein and supported by nonhydrolyzed nucleotide analogs. Maximal peptidase activity requires ATP or dATP. Replacement of Ms-Lon's catalytic Ser with Ala (S675A), Thr (S675T), or Cys (S675C) reduced to background levels Ms-Lon's in vitro peptidase activity. However, by employing a sensitive in vivo assay, based on the degradation of RcsA, we demonstrated that the S675C variant retained specific protease activity. Finally, variants of Ms-Lon, with substututions at or near S675, reduce the enzyme's basal ATPase activity, suggesting a structural interaction between the peptidase and ATPase active sites of Ms-Lon.

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“…Many genes, including the ats and mao operons regulated by monoamine compounds, have been found and classified into the monoamine (moa) regulon, which is positively regulated by moaR (1,2). The precise role of monoamine oxidase which belongs to the moa regulon is unknown either in E. coli or in K. aerogenes.…”

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confidence: 99%

maoB, a gene that encodes a positive regulator of the monoamine oxidase gene (maoA) in Escherichia coli

et al. 1996

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The structural gene for copper-and topa quinone-containing monoamine oxidase (maoA) and an unknown amine oxidase gene have been located at 30.9 min on the Escherichia coli chromosome. Deletion analysis showed that the unknown gene was located within a 1.1-kb cloned fragment adjacent to the maoA gene. The nucleotide sequence of this fragment was determined, and a single open reading frame (maoB) consisting of 903 bp was found. The gene encoded a polypeptide with a predicted molecular mass of 34,619 Da which was correlated with the migration on a sodium dodecyl sulfate-polyacrylamide gel. The predicted amino acid sequence of the MaoB protein was identical to the NH 2 -terminal amino acid sequence derived by Edman degradation of the protein synthesized under the self-promoter. No homology of the nucleotide sequence of maoB to the sequences of any reported genes was found. However, the amino acid sequence of MaoB showed a high level of homology with respect to the helix-turn-helix motif of the AraC family in its C terminus. The homology search and disruption of maoA on the chromosome led to the conclusion that MaoB is a transcriptional activator of maoA but not an amine oxidase. The consensus sequence of the cyclic AMP-cyclic AMP receptor protein complex binding domain was adjacent to the putative promoter for the maoB gene. By use of lac gene fusions with the maoA and maoB genes, we showed that the maoA gene is regulated by tyramine and MaoB and that the expression of the maoB gene is subject to catabolite repression. Thus, it seems likely that tyramine and the MaoB protein activate the transcription of maoA by binding to the regulatory region of the maoA gene.

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A Klebsiella aerogenes moaEF operon is controlled by the positive MoaR regulator of the monoamine regulon

Cited by 8 publications

References 13 publications

The Iron Sulfur Protein AtsB Is Required for Posttranslational Formation of Formylglycine in the Klebsiella Sulfatase

The Iron Sulfur Protein AtsB Is Required for Posttranslational Formation of Formylglycine in the Klebsiella Sulfatase

The Lon Protease from Mycobacterium smegmatis: Molecular Cloning, Sequence Analysis, Functional Expression, and Enzymatic Characterization

maoB, a gene that encodes a positive regulator of the monoamine oxidase gene (maoA) in Escherichia coli

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