Role of the upstream region containing an intrinsic DNA curvature in the negative regulation of the phospholipase C gene of Clostridium perfringens

T. Toyonaga, Osamu Matsushita, S. Katayama, J. Minami, A. Okabe

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The phospholipase C (α-toxin) gene (plc) of Clostridium perfringens was cloned into pUC19 and the effects of the upstream regions on expression of the plc gene were examined in Escherichia coli JM 109. When the 0.7-kb region just upstream of the putative - 35 site of the gene was deleted, production of phospholipase C increased approximately 10-fold. Northern blot hybridization analysis of the plc transcript showed that the upstream region inhibited transcription from the plc promoter. Nucleotide sequencing of this upstream region revealed that there are three periodically repeated (dA)5-6 tracts between positions -66 and -40 of the plc gene. A fragment containing this sequence showed anomalously slow electrophoretic mobility at low temperature, indicating that the region immediately upstream of the plc promoter is a locus of sequence directed DNA-bending. Nested deletions of the upstream region were created from its 5' end by exonuclease III and the effects of deletions on the expression of the plc gene were examined. When the 77-bp fragment containing the two (dA)5-6 tracts was deleted, phospholipase C production increased markedly. These results indicate that the intrinsic DNA curvature upstream of the plc promoter is involved in the negative regulation of the plc gene transcription.

Original languageEnglish
Pages (from-to)603-613
Number of pages11
JournalMicrobiology and Immunology
Volume36
Issue number6
Publication statusPublished - 1992

Fingerprint

Clostridium perfringens
Type C Phospholipases
DNA
Genes
Phosphodiesterase I
Gene Expression
Northern Blotting
Nucleotides
Escherichia coli
Temperature

ASJC Scopus subject areas

  • Immunology and Microbiology(all)
  • Microbiology
  • Microbiology (medical)

Cite this

Role of the upstream region containing an intrinsic DNA curvature in the negative regulation of the phospholipase C gene of Clostridium perfringens. / Toyonaga, T.; Matsushita, Osamu; Katayama, S.; Minami, J.; Okabe, A.

In: Microbiology and Immunology, Vol. 36, No. 6, 1992, p. 603-613.

Research output: Contribution to journalArticle

@article{d732b21ebaca4b308667692cf07ba7c0,
title = "Role of the upstream region containing an intrinsic DNA curvature in the negative regulation of the phospholipase C gene of Clostridium perfringens",
abstract = "The phospholipase C (α-toxin) gene (plc) of Clostridium perfringens was cloned into pUC19 and the effects of the upstream regions on expression of the plc gene were examined in Escherichia coli JM 109. When the 0.7-kb region just upstream of the putative - 35 site of the gene was deleted, production of phospholipase C increased approximately 10-fold. Northern blot hybridization analysis of the plc transcript showed that the upstream region inhibited transcription from the plc promoter. Nucleotide sequencing of this upstream region revealed that there are three periodically repeated (dA)5-6 tracts between positions -66 and -40 of the plc gene. A fragment containing this sequence showed anomalously slow electrophoretic mobility at low temperature, indicating that the region immediately upstream of the plc promoter is a locus of sequence directed DNA-bending. Nested deletions of the upstream region were created from its 5' end by exonuclease III and the effects of deletions on the expression of the plc gene were examined. When the 77-bp fragment containing the two (dA)5-6 tracts was deleted, phospholipase C production increased markedly. These results indicate that the intrinsic DNA curvature upstream of the plc promoter is involved in the negative regulation of the plc gene transcription.",
author = "T. Toyonaga and Osamu Matsushita and S. Katayama and J. Minami and A. Okabe",
year = "1992",
language = "English",
volume = "36",
pages = "603--613",
journal = "Microbiology and Immunology",
issn = "0385-5600",
publisher = "Center for Academic Publications Japan",
number = "6",

}

TY - JOUR

T1 - Role of the upstream region containing an intrinsic DNA curvature in the negative regulation of the phospholipase C gene of Clostridium perfringens

AU - Toyonaga, T.

AU - Matsushita, Osamu

AU - Katayama, S.

AU - Minami, J.

AU - Okabe, A.

PY - 1992

Y1 - 1992

N2 - The phospholipase C (α-toxin) gene (plc) of Clostridium perfringens was cloned into pUC19 and the effects of the upstream regions on expression of the plc gene were examined in Escherichia coli JM 109. When the 0.7-kb region just upstream of the putative - 35 site of the gene was deleted, production of phospholipase C increased approximately 10-fold. Northern blot hybridization analysis of the plc transcript showed that the upstream region inhibited transcription from the plc promoter. Nucleotide sequencing of this upstream region revealed that there are three periodically repeated (dA)5-6 tracts between positions -66 and -40 of the plc gene. A fragment containing this sequence showed anomalously slow electrophoretic mobility at low temperature, indicating that the region immediately upstream of the plc promoter is a locus of sequence directed DNA-bending. Nested deletions of the upstream region were created from its 5' end by exonuclease III and the effects of deletions on the expression of the plc gene were examined. When the 77-bp fragment containing the two (dA)5-6 tracts was deleted, phospholipase C production increased markedly. These results indicate that the intrinsic DNA curvature upstream of the plc promoter is involved in the negative regulation of the plc gene transcription.

AB - The phospholipase C (α-toxin) gene (plc) of Clostridium perfringens was cloned into pUC19 and the effects of the upstream regions on expression of the plc gene were examined in Escherichia coli JM 109. When the 0.7-kb region just upstream of the putative - 35 site of the gene was deleted, production of phospholipase C increased approximately 10-fold. Northern blot hybridization analysis of the plc transcript showed that the upstream region inhibited transcription from the plc promoter. Nucleotide sequencing of this upstream region revealed that there are three periodically repeated (dA)5-6 tracts between positions -66 and -40 of the plc gene. A fragment containing this sequence showed anomalously slow electrophoretic mobility at low temperature, indicating that the region immediately upstream of the plc promoter is a locus of sequence directed DNA-bending. Nested deletions of the upstream region were created from its 5' end by exonuclease III and the effects of deletions on the expression of the plc gene were examined. When the 77-bp fragment containing the two (dA)5-6 tracts was deleted, phospholipase C production increased markedly. These results indicate that the intrinsic DNA curvature upstream of the plc promoter is involved in the negative regulation of the plc gene transcription.

UR - http://www.scopus.com/inward/record.url?scp=0026748951&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026748951&partnerID=8YFLogxK

M3 - Article

C2 - 1522810

AN - SCOPUS:0026748951

VL - 36

SP - 603

EP - 613

JO - Microbiology and Immunology

JF - Microbiology and Immunology

SN - 0385-5600

IS - 6

ER -