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Characterization of Chlamydomonas reinhardtii Zygote-Specific cDNAs That Encode Novel Proteins Containing Ankyrin Repeats and WW Domains1

Hideo Kuriyama*, Hiroyoshi Takano, Lena Suzuki, Hidenobu Uchida, Shigeyuki Kawano, Haruko Kuroiwa, and Tsuneyoshi Kuroiwa

Department of Biological Sciences, Graduate School of Science, University of Tokyo, Hongo, Tokyo 113, Japan (H.K., H.T., S.K., T.K.); Department of Hygiene and Oncology, Graduate School of Medicine, Tokyo Medical and Dental University, Yushima, Tokyo 113, Japan (L.S.); Institute of Biological Science, University of Tsukuba, Tsukuba, Ibaraki 305, Japan (H.U.); and Department of Biology, Kyoritsu Women's Junior College, Tokyo 102, Japan (H. Kuroiwa)

Genes that are expressed only in the young zygote are considered to be of great importance in the development of an isogamous green alga, Chlamydomonas reinhardtii. Clones representing the Zys3 gene were isolated from a cDNA library prepared using zygotes at 10 min after fertilization. Sequencing of Zys3 cDNA clones resulted in the isolation of two related molecular species. One of them encoded a protein that contained two kinds of protein-to-protein interaction motifs known as ankyrin repeats and WW domains. The other clone lacked the ankyrin repeats but was otherwise identical. These mRNA species began to accumulate simultaneously in cells beginning 10 min after fertilization, and reached maximum levels at about 4 h, after which time levels decreased markedly. Genomic DNA gel-blot analysis indicated that Zys3 was a single-copy gene. The Zys3 proteins exhibited parallel expression to the Zys3 mRNAs at first, appearing 2 h after mating, and reached maximum levels at more than 6 h, but persisted to at least 1 d. Immunocytochemical analysis revealed their localization in the endoplasmic reticulum, which suggests a role in the morphological changes of the endoplasmic reticulum or in the synthesis and transport of proteins to the Golgi apparatus or related vesicles.

1   This work was supported by a Grant-in-Aid for Special Promoted Research (project no. 06101002 to T.K.) from the Ministry of Education, Science and Culture of Japan.
*   Corresponding author; e-mail kuriyam2{at}gol.com or kuriyama{at}biol.s.u-tokyo.ac.jp; fax 81-3-3812-4929.

Plant Physiol. (1999) 119: 873-884
Copyright Clearance Center:   0032-0889/99/119//12
© 1999 American Society of Plant Physiologists




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