| Mutations in DnaA protein suppress the growth arrest of acidic phospholipid-deficient Escherichia coli cells. | |
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MedLine Citation:
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PMID: 11230139 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Cell growth arrests when the concentrations of anionic phospholipids drop below a critical level in Escherichia coli, with the insufficient amounts of acidic phospholipids adversely affecting the DnaA-dependent initiation of DNA replication at the chromosomal origin (oriC). Mutations have been introduced into the carboxyl region of DnaA, including the portion identified as essential for productive in vitro DnaA-acidic phospholipid interactions. Expression of DnaA proteins possessing certain small deletions or substituted amino acids restored growth to cells deficient in acidic phospholipids, whereas expression of wild-type DnaA did not. The mutations include substitutions and deletions in the phospholipid-interacting domain as well as some small deletions in the DNA-binding domain of DnaA. Marker frequency analysis indicated that initiation of replication occurs at or near oriC in acidic phospholipid- deficient cells rescued by the expression of DnaA having a point mutation in the membrane-binding domain, DnaA(L366K). Flow cytometry revealed that expression in wild-type cells of plasmid-borne DnaA(L366K) and DnaA(Delta363-367) reduced the frequency with which replication was initiated and disturbed the synchrony of initiations. |
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Authors:
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W Zheng; Z Li; K Skarstad; E Crooke |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: The EMBO journal Volume: 20 ISSN: 0261-4189 ISO Abbreviation: EMBO J. Publication Date: 2001 Mar |
Date Detail:
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Created Date: 2001-03-20 Completed Date: 2001-04-19 Revised Date: 2009-11-18 |
Medline Journal Info:
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Nlm Unique ID: 8208664 Medline TA: EMBO J Country: England |
Other Details:
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Languages: eng Pagination: 1164-72 Citation Subset: IM |
Affiliation:
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Department of Biochemistry and Molecular Biology, Georgetown University Medical Center, 3900 Reservoir Road NW, Washington DC 20007, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Substitution
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genetics Bacterial Proteins / analysis, chemistry, genetics*, metabolism* Cell Division Cephalexin / pharmacology Chromosomes, Bacterial / genetics DNA Replication / drug effects DNA, Bacterial / biosynthesis, genetics DNA-Binding Proteins / chemistry, genetics*, metabolism* Escherichia coli / cytology*, genetics, growth & development, metabolism Flow Cytometry Hydrogen-Ion Concentration Origin Recognition Complex Phospholipids / chemistry, deficiency* Plasmids / genetics Protein Structure, Tertiary Rifampin / pharmacology Sequence Deletion / genetics Suppression, Genetic / genetics* Transformation, Bacterial Viral Proteins / genetics |
| Grant Support | |
ID/Acronym/Agency:
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GM49700/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Bacterial Proteins; 0/DNA, Bacterial; 0/DNA-Binding Proteins; 0/DnaA protein, Bacteria; 0/OriC chromosomal replication origin; 0/Origin Recognition Complex; 0/Phospholipids; 0/Viral Proteins; 13292-46-1/Rifampin; 15686-71-2/Cephalexin |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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