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Plant and Cell Physiology Advance Access originally published online on January 28, 2009
Plant and Cell Physiology 2009 50(3):572-583; doi:10.1093/pcp/pcp016
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

The Chlamydomonas Hatching Enzyme, Sporangin, is Expressed in Specific Phases of the Cell Cycle and is Localized to the Flagella of Daughter Cells Within the Sporangial Cell Wall

Takeaki Kubo1,2,*, Shinsuke Kaida3, Jun Abe3,4, Tatsuaki Saito2, Hideya Fukuzawa1 and Yoshihiro Matsuda3

1Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kyoto, 606-8502 Japan
2Department of Applied Science, Faculty of Science, Okayama University of Science, Okayama, 700-0005 Japan
3Department of Biology, Faculty of Science, Kobe University, Kobe, 657-8501 Japan

*Corresponding author: E-mail, kubot{at}lif.kyoto-u.ac.jp; Fax, +81-75-753-6127.


  Abstract

The timely breakdown of the extracellular matrix by proteolytic enzymes is essential for development, morphogenesis and cell proliferation in plant and animal cells. Sporangin of the unicellular green alga Chlamydomonas reinhardtii that mediates breakdown of the sporangial cell wall to liberate the daughter cells after cell division is characterized as a subtilase-like serine protease. The sporangin gene is specifically transcribed during S/M phase in a synchronized vegetative cell cycle. In immunoblot analyses using a polyclonal antibody raised against the sporangin polypeptide, the enzyme is synthesized after mitotic cell division and accumulated in the daughter cells before hatching. Immunofluorescence analyses showed that sporangin is localized to the flagella of the daughter cells within the sporangial cell wall, and released into the culture medium. The data suggest that sporangin is released from flagella concurrently with the digestion of sporangial cell wall, and then the daughter cells are hatched from the sporangia in the Chlamydomonas vegetative cell cycle.

Keywords: Cell cycle - Chlamydomonas reinhardtii - Flagella - Hatching enzyme - Subtilase-like serine protease.

Abbreviations: ECM, extracellular matrix; EST, expressed sequence tag; GST, glutathione S-transferase; PBS, phosphate-buffered saline; RACE, rapid amplification of cDNA ends

4Present address: Department of Chemical and Biological Sciences, Faculty of Science, Japan Woman's University, Tokyo, 112-8681 Japan

(Received December 23, 2008; Accepted January 25, 2009)
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