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Vol. 10, Issue 5, 1637-1652, May 1999

Expression and Differential Intracellular Localization of Two Major Forms of Human 8-Oxoguanine DNA Glycosylase Encoded by Alternatively Spliced OGG1 mRNAs

Kenichi Nishioka,* Toshio Ohtsubo,* Hisanobu Oda,* Toshiyuki Fujiwara,dagger Dongchon Kang,Dagger Keizo Sugimachi,§ and Yusaku Nakabeppu*parallel

 *Department of Biochemistry, Medical Institute of Bioregulation,  Dagger Department of Clinical Chemistry and Laboratory Medicine, Faculty of Medicine,  §Department of Surgery, Faculty of Medicine, Kyushu University, Fukuoka 812-8582, Japan; and  dagger Department of Biochemistry, School of Medicine, Fukuoka University, Fukuoka 814-0180, Japan

We identified seven alternatively spliced forms of human 8-oxoguanine DNA glycosylase (OGG1) mRNAs, classified into two types based on their last exons (type 1 with exon 7: 1a and 1b; type 2 with exon 8: 2a to 2e). Types 1a and 2a mRNAs are major in human tissues. Seven mRNAs are expected to encode different polypeptides (OGG1-1a to 2e) that share their N terminus with the common mitochondrial targeting signal, and each possesses a unique C terminus. A 36-kDa polypeptide, corresponding to OGG1-1a recognized only by antibodies against the region containing helix-hairpin-helix-PVD motif, was copurified from the nuclear extract with an activity introducing a nick into DNA containing 8-oxoguanine. A 40-kDa polypeptide corresponding to a processed form of OGG1-2a was detected in their mitochondria using antibodies against its C terminus. Electron microscopic immunocytochemistry and subfractionation of the mitochondria revealed that OGG1-2a locates on the inner membrane of mitochondria. Deletion mutant analyses revealed that the unique C terminus of OGG1-2a and its mitochondrial targeting signal are essential for mitochondrial localization and that nuclear localization of OGG1-1a depends on the NLS at its C terminus.


parallel    Corresponding author. E-mail address: yusaku{at}bioreg.kyushu-u.ac.jp.


Molecular Biology of the Cell
Vol. 10, 1637-1652, May 1999
Copyright © 1999 by The American Society for Cell Biology



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