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PMID: 2786875 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The 44P subunit of the T4 DNA polymerase accessory protein complex catalyzes ATP hydrolysis.

The Journal of biological chemistry ·Vol. 264 ·No. 19 ·1989-07-05 ·Pages 10943-53

Rush J, Lin TC, Quinones M, Spicer EK, Douglas I, Williams KR, Konigsberg WH

Abstract

The genes encoding all three T4 DNA polymerase accessory proteins have been cloned into overexpression plasmids. Induction of cells harboring these plasmids results in the synthesis of each accessory protein at levels that approach 10% of the total cellular protein. The solubility of the accessory proteins after induction at 42 degrees C ranges from about 60% to greater than 95%. A plasmid that allows overexpression of the 44P/62P complex has been manipulated further to overexpress selectively the 44P subunit without 62P, permitting us to assess how each subunit contributes to the properties of the 44P/62P complex. A comparison of 44P and 44P/62P by conventional hydrodynamic techniques shows that 44P forms a subcomplex nearly as large as the 44P/62P complex. In addition, 44P catalyzes DNA-dependent ATP hydrolysis with a specific activity similar to that of the 44P/62P ATPase. However, unlike the 44P/62P complex, the ATPase activity of 44P alone is only slightly stimulated by 45P. This suggests that one role of the 62P subunit is to facilitate a productive interaction of 44P and 45P.

MeSH Terms
Adenosine Triphosphate/metabolism Amino Acid Sequence Base Sequence Chromatography, DEAE-Cellulose Chromatography, Gel Cloning, Molecular Codon Gene Expression Regulation Genes, Viral Hydrolysis Molecular Sequence Data Molecular Weight Plasmids T-Phages/genetics Transformation, Genetic Ultracentrifugation Viral Proteins/genetics,isolation & purification,metabolism
Chemicals
Codon Viral Proteins Adenosine Triphosphate
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Rush J
Department of Molecular Biophysics and Biochemistry, Yale School of Medicine, New Haven, Connecticut 06510.
Lin T C
Quinones M
Spicer E K
Douglas I
Williams K R
Konigsberg W H
Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
Published
1989-07-05
Pages
10943-53
Language
English
Region
United States
NLM ID
2985121R
Subset
IM
Grants
NIGMS NIH HHS · GM12601-20 · United States
NIGMS NIH HHS · GM30191 · United States
Databases
GENBANK
J02510, M10160, X00769
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