Characterization of a Bacillus subtilis 64-kDa DNA polymerase X potentially involved in DNA repair.

Abstract:

Bacillus subtilis gene yshC encodes a 64-kDa family X DNA polymerase (PolXBs), which contains all the critical residues involved in DNA and nucleotide binding as well as those responsible for catalysis of DNA polymerization, conserved in most family X members. Biochemical analyses of the purified enzyme indicate that PolXBs is a monomeric and strictly template-directed DNA polymerase, preferentially acting on DNA structures containing gaps from one to a few nucleotides and bearing a phosphate group at the 5' end of the downstream DNA. The fact that PolXBs is able to conduct filling of a single-nucleotide gap, allowing further sealing of the resulting nick by a DNA ligase, points to a putative role in base excision repair during the B. subtilis life cycle.

Polymerases:

Topics:

Reverse Transcriptase, Terminal Transferase, Structure and Structure/Function, Exonuclease Activity, Source / Purification, Other Enzymatic Activities, RNase H Activity, Nucleotide Incorporation

One line summary:

Biochemical characterization of Bacillus subtilis DNA polymerase X as a potential DNA repair enzyme

Status:

new topics/pols set partial results complete validated

Results:

Polymerase Reference Property Result Context
Bsu Pol X Baños B2008 Reverse Transcriptase Activity No
Bsu Pol X Baños B2008 Terminal Transferase No
Bsu Pol X Baños B2008 Polymerase Catalytic Residue Amino Acids D193, D195, D240
Bsu Pol X Baños B2008 3-5' Exonuclease (proofreading) Yes
Bsu Pol X Baños B2008 Cloned or native Cloned in E. coli
Bsu Pol X Baños B2008 Residues Involved in Catalysis of 3-5' Exo H339, H341, H437, H465, H528, E410, E496 and D526
Bsu Pol X Baños B2008 5-3' Exonuclease No
Bsu Pol X Baños B2008 Tagged Yes
Bsu Pol X Baños B2008 Tag Name His-Tag
Bsu Pol X Baños B2008 Full length or truncated Full length
Bsu Pol X Baños B2008 Extension from RNA primer Unspecified
Bsu Pol X Baños B2008 RNase H No
Bsu Pol X Baños B2008 Nick Extension No
Bsu Pol X Baños B2008 Gap Filling Yes

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