title: Cloning, expression, purification, crystallization and preliminary crystallographic analysis of MsDpo4: a Y-family DNA polymerase from Mycobacterium smegmatis.
abstract: The expression of error-prone DNA polymerases belonging to the Y-family is upregulated in prokaryotes under adverse conditions to facilitate adaptive mutagenesis. However, it has been suggested that representatives of this family in mycobacteria do not participate in adaptive mutagenesis. These studies raise the possibility that mycobacterial representatives might be devoid of biochemical activity. In order to determine whether this possible loss of activity is a consequence of significant changes in the structure of these enzymes, structural studies on a representative enzyme from Mycobacterium smegmatis, MsDpo4, were initiated. The protein crystallized in space group P6(1)22 or P6(5)22. The Matthews coefficient was 4.0 Å3 Da(-1) assuming the presence of one molecule in the asymmetric unit; the corresponding solvent content was 69%. X-ray diffraction data were collected to a minimum Bragg spacing of 2.6 Å and crystals of selenomethionine-labelled MsDpo4 have been prepared for ab initio phasing.
authors.affiliation: National Centre for Biological Sciences (NCBS-TIFR), UAS GKVK Campus, Bellary Road, Bangalore 560 065, India.
authors.forename: Amit
authors.initials: A
authors.lastname: Sharma
authors.forename: Deepak T
authors.initials: DT
authors.lastname: Nair
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry*; D015870:Gene Expression; D008969:Molecular Sequence Data; D020102:Mycobacterium smegmatis / Q000201:enzymology* / Q000302:isolation & purification; D016415:Sequence Alignment
chemical_list: D004259:DNA-Directed DNA Polymerase