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The Helicobacter pylori gene encoding phosphatidylserine synthase: sequence, expression, and insertional mutagenesis
1Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Canada.
Journal of Bacteriology
|August 1, 1997
Summary
The Helicobacter pylori pss gene encodes phosphatidylserine synthase (PSS). This enzyme is essential for H. pylori growth, as attempts to disrupt the gene were unsuccessful, indicating its vital role.
Area of Science:
- Microbiology
- Molecular Biology
- Biochemistry
Background:
- Helicobacter pylori is a significant human pathogen.
- Phosphatidylserine synthase (PSS) is crucial for cell membrane biosynthesis.
- Understanding PSS in H. pylori can reveal novel therapeutic targets.
Purpose of the Study:
- To clone and characterize the H. pylori pss gene.
- To investigate the function and essentiality of PSS in H. pylori.
Main Methods:
- Gene cloning and sequencing of H. pylori pss.
- Expression analysis in E. coli maxicells.
- Enzyme activity assays on membrane fractions.
- Gene disruption attempts using a chloramphenicol acetyltransferase cassette.
Main Results:
- The H. pylori pss gene encodes a 237-amino acid polypeptide.
- H. pylori PSS shares sequence homology with PSS from various organisms but not E. coli.
- Expressed H. pylori PSS exhibits manganese-dependent activity and is membrane-bound.
- Disruption of the pss gene in H. pylori UA802 was unsuccessful.
Conclusions:
- The H. pylori pss gene product is a functional, membrane-bound enzyme.
- Phosphatidylserine synthase is essential for the growth of H. pylori.
- The essentiality of PSS suggests it as a potential target for anti-H. pylori therapies.