scholarly journals Purification of the rep protein of Escherichia coli. An ATPase which separates duplex DNA strands in advance of replication.

1978 ◽  
Vol 253 (9) ◽  
pp. 3292-3297 ◽  
Author(s):  
J.F. Scott ◽  
A. Kornberg
Keyword(s):  
Escherichia Coli ◽  
Duplex Dna ◽  
Rep Protein ◽  
Dna Strands

The rep mutation. VI. Purification and properties of the Escherichia coli rep protein, DNA helicase III

1979 ◽  
Vol 57 (6) ◽  
pp. 855-866 ◽  
Author(s):  
Seishi Takahashi ◽  
Christian Hours ◽  
Alan Chu ◽  
David T. Denhardt
Keyword(s):  
Escherichia Coli ◽  
Cell Extract ◽  
Dna Helicase ◽  
Protein Product ◽  
Duplex Dna ◽  
Replicative Form ◽  
Single Stranded Dna ◽  
Rep Protein ◽  
E Coli

The protein product of the rep gene of Escherichia coli is required for the replication of certain bacteriophage genomes ([Formula: see text], fd, P2) and for the normal replication of E. coli DNA. We have used a specialized transducing phage, λp rep+, which complements the defect of rep mutants, to identify the rep protein. The rep protein has been purified from cells infected with λp rep+ phage; it has a molecular weight of about 70 000 and appears similar to the protein found in normal cells. Stimulation of [Formula: see text] replicative form DNA synthesis in vitro was observed when highly purified rep protein was supplied to a cell extract derived from [Formula: see text]-infected E. coli rep cells and supplemented with replicative form DNA. The purified protein has a single-stranded DNA-dependent ATPase activity and is capable of sensitizing duplex DNA to nucleases specific for single-stranded DNA. For this reason we propose the enzyme be called DNA helicase III. We infer that the rep protein uses the energy of hydrolysis of ATP to separate the strands of duplex DNA; the E. coli DNA binding protein need not be present. The rep3 mutant appeared to make a limited amount of active rep protein.

scholarly journals ATP utilization by rep protein in the catalytic separation of DNA strands at a replicating fork.

1978 ◽  
Vol 253 (9) ◽  
pp. 3298-3304 ◽  
Author(s):  
A. Kornberg ◽  
J.F. Scott ◽  
L.L. Bertsch
Keyword(s):  
Rep Protein ◽  
Dna Strands
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