Interaction of apurinic/apyrimidinic endonuclease 2 (Apn2) with Myh1 DNA glycosylase in fission yeast

详细信息    查看全文

• 作者：Jin Jin^a ; Bor-Jang Hwang^a ; Po-Wen Chang^a ; Eric A. Toth^a ; ^b ; ^c ; A-Lien Lu^a ; ^b ; ^{aluchang@umaryland.edu" class="auth_mail}
• 关键词：3&prime ; -dRP ; 3&prime ; -伪 ; 尾-unsaturated aldehyde ; 8-oxoG or G° ; 7 ; 8-dihydro-8-oxoguanine ; 9&ndash ; 1&ndash ; 1 ; Rad9&ndash ; Rad1&ndash ; Hus1 ; AP ; apurinic/apyrimidinic ; APE1 or Apn1 ; AP-endonuclease 1 ; Apn2 ; AP-endonuclease 2 ; BER ; base excision repair ; BSA ; bovine serum albumin ; DTT ; dithiothreitol ; FAM ; fluorescein ; GST ; glutathione S-transferase ; h ; human ; IDC ; interdomain connector ; kobs ; rate constants ; MAP ; MYH-associated polyposis ; MBP ; maltose binding protein ; MYH ; MUTYH ; or Myh1 ; MutY h
• 刊名：DNA Repair
• 出版年：March, 2014
• 年：2014
• 卷：15
• 期：Complete
• 页码：1-10
• 全文大小：2347 K

文摘

Oxidative DNA damage is repaired primarily by the base excision repair (BER) pathway in a process initiated by removal of base lesions or mismatched bases by DNA glycosylases. MutY homolog (MYH, MUTYH, or Myh1) is a DNA glycosylase which excises adenine paired with the oxidative lesion 8-oxo-7,8-dihydroguanine (8-oxoG, or G掳), thus reducing G:C to T:A mutations. The resulting apurinic/apyrimidinic (AP) site is processed by an AP-endonuclease or a bifunctional glycosylase/lyase. We show here that the major Schizosaccharomyces pombe AP endonuclease, Apn2, binds to the inter-domain connector located between the N- and C-terminal domains of Myh1. This Myh1 inter-domain connector also interacts with the Hus1 subunit of the Rad9-Rad1-Hus1 checkpoint clamp. Mutagenesis studies indicate that Apn2 and Hus1 bind overlapping but different sequence motifs on Myh1. Mutation on I²⁶¹ of Myh1 reduces its interaction with Hus1, but only slightly attenuates its interaction with Apn2. However, E²⁶² of Myh1 is a key determinant for both Apn2 and Hus1 interactions. Like human APE1, Apn2 has 3鈥?phosphodiesterase activity. However, unlike hAPE1, Apn2 has a weak AP endonuclease activity which cleaves the AP sites generated by Myh1 glycosylase. Functionally, Apn2 stimulates Myh1 glycosylase activity and Apn2 phosphodiesterase activity is stimulated by Myh1. The cross stimulation of Myh1 and Apn2 enzymatic activities is dependent on their physical interaction. Thus, Myh1 and Apn2 constitute an initial BER complex.