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Isolation, expression, and nucleotide sequence of the sod gene from Porphyromonas gingivalis.

ABSTRACT: The sod gene coding for the Mn/Fe-dependent superoxide dismutase (SOD) enzyme has been isolated on a 5.9-kb DNA fragment from Porphyromonas gingivalis ATCC 53977. SOD activity can be expressed from the P. gingivalis fragment and from a subcloned fragment in Escherichia coli. However, the enzyme does not appear to be expressed from its own promoter in E. coli cells. The nucleotide sequence of the gene has been determined, and the deduced amino acid sequence of the enzyme is nearly identical to that of the enzyme purified from P. gingivalis 381 and shares extensive sequence similarity with comparable enzymes from E. coli.

SUBMITTER: Choi JI

PROVIDER: S-EPMC257877 | biostudies-other | 1991 Apr

REPOSITORIES: biostudies-other

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Isolation, expression, and nucleotide sequence of the sod gene from Porphyromonas gingivalis.

Choi J I JI Takahashi N N Kato T T Kuramitsu H K HK

Infection and immunity 19910401 4

The sod gene coding for the Mn/Fe-dependent superoxide dismutase (SOD) enzyme has been isolated on a 5.9-kb DNA fragment from Porphyromonas gingivalis ATCC 53977. SOD activity can be expressed from the P. gingivalis fragment and from a subcloned fragment in Escherichia coli. However, the enzyme does not appear to be expressed from its own promoter in E. coli cells. The nucleotide sequence of the gene has been determined, and the deduced amino acid sequence of the enzyme is nearly identical to th ...[more]

PMID: 1840572

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Project description:We have previously reported on a Tn4351-generated mutant of Porphyromonas gingivalis (MSM-3) which expresses enhanced arginine-specific proteinase activity and does not utilize hemin or hemoglobin for growth (C. A. Genco et al., Infect. Immun. 63:2459-2466, 1995). In the process of characterizing the genetic lesion in P. gingivalis MSM-3, we have determined that the endogenous P. gingivalis insertion sequence element IS1126 is capable of transposition within P. gingivalis. We have also determined that IS1126 transposition modulates the transcription of the genes encoding the lysine-specific proteinase, gingipain K (kgp) and the arginine-specific proteinase, gingipain R2 (rgpB). Sequence analysis of P. gingivalis MSM-3 revealed that Tn4351 had inserted 60 bp upstream of the P. gingivalis endogenous IS element IS1126. Furthermore, P. gingivalis MSM-3 exhibited two additional copies of IS1126 compared to the parental strain A7436. Examination of the first additional IS1126 element, IS1126(1), indicated that it has inserted into the putative promoter region of the P. gingivalis kgp gene. Analysis of total RNA extracted from P. gingivalis MSM-3 demonstrated no detectable kgp transcript; likewise, P. gingivalis MSM-3 was devoid of lysine-specific proteinase activity. The increased arginine-specific proteinase activity exhibited by P. gingivalis MSM-3 was demonstrated to correlate with an increase in the rgpA and rgpB transcripts. The second additional IS1126 element, IS1126(2), was found to have inserted upstream of a newly identified gene, hmuR, which exhibits homology to a number of TonB-dependent genes involved in hemin and iron acquisition. Analysis of total RNA from P. gingivalis MSM-3 demonstrated that hmuR is transcribed, indicating that the insertion of IS1126 had not produced a polar effect on hmuR transcription. The hemin-hemoglobin defect in P. gingivalis MSM-3 is proposed to result from the inactivation of Kgp, which has recently been demonstrated to function in hemoglobin binding. Taken together, the results presented here demonstrate that the introduction of Tn4351 into the P. gingivalis chromosome has resulted in two previously undocumented phenomena in P. gingivalis: (i) the transposition of the endogenous insertion sequence element IS1126 and (ii) the modulation of gingipain transcription and translation as a result of IS1126 transposition.

Dataset Information

Isolation, expression, and nucleotide sequence of the sod gene from Porphyromonas gingivalis.

Publications

Isolation, expression, and nucleotide sequence of the sod gene from Porphyromonas gingivalis.

Similar Datasets

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