Document Detail


Restoration of growth to acidic phospholipid-deficient cells by DnaA(L366K) is independent of its capacity for nucleotide binding and exchange and requires DnaA.
MedLine Citation:
PMID:  15642730     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In the absence of adequate levels of cellular acidic phospholipids, Escherichia coli remain viable but are arrested for growth. Expression of a DnaA protein that contains a single amino acid substitution in the membrane-binding domain, DnaA(L366K), in concert with expression of wild-type DnaA protein, restores growth. DnaA protein has high affinity for ATP and ADP, and in vitro lipid bilayers that are fluid and contain acidic phospholipids reactivate inert ADP-DnaA by promoting an exchange of ATP for ADP. Here, nucleotide and lipid interactions and replication activity of purified DnaA(L366K) were examined to gain insight into the mechanism of how it restores growth to cells lacking acidic phospholipids. DnaA(L366K) behaved like wild-type DnaA with respect to nucleotide binding affinities and hydrolysis properties, specificity of acidic phospholipids for nucleotide release, and origin binding. Yet, DnaA(L366K) was feeble at initiating replication from oriC unless augmented with a limiting quantity of wild-type DnaA, reflecting the in vivo requirement that both wild-type and a mutant form of DnaA must be expressed and act together to restore growth to acidic phospholipid deficient cells.
Authors:
Zhenya Li; Jennifer L Kitchen; Kelly Boeneman; Priyanka Anand; Elliott Crooke
Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.     Date:  2005-01-10
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  280     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2005 Mar 
Date Detail:
Created Date:  2005-03-14     Completed Date:  2005-04-25     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  9796-801     Citation Subset:  IM    
Affiliation:
Department of Biochemistry and Molecular Biology, Georgetown University Medical Center, 3900 Reservoir Rd. NW, Washington, DC 20007, USA.
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MeSH Terms
Descriptor/Qualifier:
Adenosine Diphosphate / chemistry
Adenosine Triphosphate / chemistry
Bacterial Proteins / genetics,  metabolism*
Cell-Free System
Chromosomes / metabolism
Chromosomes, Bacterial
DNA Replication
DNA-Binding Proteins / genetics,  metabolism*
Dose-Response Relationship, Drug
Escherichia coli / metabolism*
Hydrolysis
Lipid Bilayers / chemistry
Lipid Metabolism
Lipids / chemistry
Mutation
Phospholipids / chemistry,  metabolism
Protein Binding
Protein Structure, Tertiary
Replication Origin
Time Factors
Grant Support
ID/Acronym/Agency:
R01GM49700/GM/NIGMS NIH HHS; T32CA009686/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Bacterial Proteins; 0/DNA-Binding Proteins; 0/DnaA protein, Bacteria; 0/Lipid Bilayers; 0/Lipids; 0/Phospholipids; 56-65-5/Adenosine Triphosphate; 58-64-0/Adenosine Diphosphate

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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