Partial Purification of Pde1 from Saccharomyces cerevisiae: Enzymatic Redundancy for the Repair of 3'-Terminal DNA Lesions and Abasic Sites in Yeast

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• 作者：Miriam Sander and and Dindial Ramotar
• 刊名：Biochemistry
• 出版年：1997
• 出版时间：May 20, 1997
• 年：1997
• 卷：36
• 期：20
• 页码：6100 - 6106
• 全文大小：171K
• 年卷期：v.36,no.20(May 20, 1997)
• ISSN：1520-4995

文摘

Earlier work indicates that the major DNA repairphosphodiesterase (PDE) in yeast cells isthe well-characterized Apn1 protein. Apn1 demonstrates bothMg²⁺-independent PDE activity andMg²⁺-independent class II apurinic/apyrimidinic (AP) endonuclease activityand represents greater than 90% ofthe activity detected in crude extracts from wild-type yeast cells.Apn1 is related to Echerichia coliendonuclease IV, both in its enzymatic properties and its amino acidsequence. In this work, we reportthe partial purification of a novel yeast protein, Pde1, present inApn1-deficient cells. Pde1 is purified bysequential BioRex-70, PBE118, and MonoS chromatography steps using asensitive and highly specific3'-phosphoglycolate-terminated oligonucleotide-based assay as a measureof PDE activity. Mg²⁺-stimulatedPDE and Mg²⁺-stimulated class II AP endonuclease copurifyduring this procedure. These results indicatethat yeast, like many other organisms studied to date, has enzymaticredundancy for the repair of 3'-blocking groups and abasic sites.