Archives of Microbiology

, Volume 173, Issue 5, pp 412–417

phrA, the major photoreactivating factor in the cyanobacterium Synechocystis sp. strain PCC 6803 codes for a cyclobutane-pyrimidine-dimer-specific DNA photolyase

  • Wing-On Ng
  • Rodolfo Zentella
  • Yinsheng Wang
  • John-Stephen A. Taylor
  • Himadri B. Pakrasi
Original Paper

DOI: 10.1007/s002030000164

Cite this article as:
Ng, WO., Zentella, R., Wang, Y. et al. Arch. Microbiol. (2000) 173: 412. doi:10.1007/s002030000164

Abstract.

A new broad-host-range plasmid, pSL1211, was constructed for the over-expression of genes in Synechocystis sp. strain PCC 6803. The plasmid was derived from RSF1010 and an Escherichia coli over-expression plasmid, pTrcHisC. Over-expressed protein is made with a removable N-terminal histidine tag. The plasmid was used to over-express the phrA gene and purify the gene product from Synechocystis sp. strain PCC 6803. PhrA is the major ultraviolet-light-resistant factor in the cyanobacterium. The purified PhrA protein exhibited an optical absorption spectrum similar to that of the cyclobutane pyrimidine dimer (CPD) DNA photolyase from Synechococcus sp. strain PCC 6301 (Anacystis nidulans). Mass spectrometry analysis of PhrA indicated that the protein contains 8-hydroxy-5-deazariboflavin and flavin adenine dinucleotide (FADH2) as cofactors. PhrA repairs only cyclobutane pyrimidine dimer but not pyrimidine (6-4) pyrimidinone photoproducts. On the basis of these results, the PhrA protein is classified as a class I, HDF-type, CPD DNA photolyase.

Synechocystis sp. strain PCC 6803 phrA DNA photolyase Cyclobutane pyrimidine dimer Over-expression plasmid pSL1211

Copyright information

© Springer-Verlag 2000

Authors and Affiliations

  • Wing-On Ng
    • 1
  • Rodolfo Zentella
    • 1
  • Yinsheng Wang
    • 2
  • John-Stephen A. Taylor
    • 2
  • Himadri B. Pakrasi
    • 1
  1. 1.Department of Biology, Campus Box 1137, Washington University, One Brookings Dr., St. Louis, MO 63130, USAUSA
  2. 2.Department of Chemistry, Campus Box 1134, Washington University, One Brookings Dr., St. Louis, MO 63130, USAUSA