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J. Biochem, 2003, Vol. 133, No. 5 625-631
© 2003 Japanese Biochemical Society

CELL

Expression of Tom34 Splicing Isoforms in Mouse Testis and Knockout of Tom34 in Mice

Kazutoyo Terada1,+, Shota Ueno1,2, Kentaro Yomogida3, Tomoaki Imai3, Hiroshi Kiyonari4, Naoki Takeda5, Masato Yano1, Shinichi Abe2, Shinichi Aizawa4 and Masataka Mori1

1 Department of Molecular Genetics, Kumamoto University School of Medicine, Honjo 2-2-1, Kumamoto 860-0811; 2 Department of Materials and Life Science, Graduate School of Science and Technology, Kumamoto University, Kurokami 2-39-1, Kumamoto 860-8555; 3 Department of Science for Laboratory Animal Experimentation, Research Institute for Microbial Diseases, Osaka University, Yamadaoka 3-1, Suita, Osaka 565-0871; 4 Laboratory for Animal Resources and Genetic Engineering, RIKEN Center for Developmental Biology, Minatojima-minamimachi 2-2-3, Chuo-ku, Kobe, Hyogo 650-0047; and 5 Division of Transgenic Technology, Center for Animal Resources and Development (CARD), Kumamoto University, Honjo 2-2-1, Kumamoto 860-0811

The 34-kDa translocase of the outer mitochondrial membrane (Tom34) is a putative mammalian-specific factor involved in protein import into mitochondria. We analyzed the genomic sequence of the mouse Tom34 gene and found it has two alternative initial exons. Using reverse transcription and the polymerase chain reaction (RT-PCR), we found that these two mRNAs differs only in the 5'-proximal sequences corresponding to the two initial exons (exon 1a and 1b). Tom34 mRNA with exon 1a (Tom34a) is expressed ubiquitously, while that with exon 1b (Tom34b) is expressed only in mature testicular germ cells. To explore the in vivo function of Tom34 proteins, we generated Tom34-deficient mice by targeted disruption. The Tom34–/– mice were viable and grew normally and had a normal Mendelian inheritance pattern. Male as well as female Tom34–/– mice were fertile. In vitro-preprotein import into isolated mitochondria showed no apparent difference between Tom34–/– and wild-type mice. These results indicate that Tom34 is dispensable for mouse growth and development under optimal conditions.

+ To whom correspondence should be addressed. Tel: +81-96-373-5143, Fax: +81-96-373-5145, E-mail: terada{at}gpo.kumamoto-u.ac.jp


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