DNA-dependent RNA polymerase II from Candida species is a multiple zinc-containing metalloenzyme.
We have purified DNA-dependent RNA polymerase II from Candida albicans, a human pathogenic yeast. The enzyme consists of 9 polypeptides that are unique to C. albicans, their mobility on SDS-PAGE being different from the mobility of the corresponding subunits of RNA polymerase II from Saccharomyces cerevisiae or C. utilis. In the present study we also demonstrate that RNA pol II from C. albican and C. utilis are metalloproteins containing approximately 5 mol of zinc per mole of enzyme. Although prolonged dialysis in 10 or 20 mM EDTA failed to remove Zn(II) from the C. albicans enzyme, in the C. utilis enzyme 3 Zn(II) ions could be removed and then reconstituted in the presence of excess Zn(II). o-Phenanthroline (5 mM) removed Zn(II) from C. albicans enzyme irreversibly in a time-dependent fashion with concomitant loss of enzyme activity. Circular dichroism studies revealed structural changes on removal of zinc, thus suggesting a role for Zn in maintenance of structural stability. Further, we demonstrate that the largest subunit of the C. utilis enzyme and the 3 large subunits of the C. albicans enzyme can bind radioactive zinc.
Patturajan
M
M
Center for Cellular and Molecular Biology, Hyderabad, India. meera@welchlink.welch.jhu.edu
Sevugan
M
M
Chatterji
D
D
D001665:Binding Sites; D002175:Candida / Q000201:enzymology*; D002176:Candida albicans / Q000201:enzymology; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006801:Humans; D008970:Molecular Weight; D020836:Protein Structure, Quaternary; D012319:RNA Polymerase II / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D015032:Zinc / Q000737:chemistry
D012319:RNA Polymerase II; D015032:Zinc