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Triple Immunofluorescent Labeling of FtsZ, Dynamin, and EF-Tu Reveals a Loose Association Between the Inner and Outer Membrane Mitochondrial Division Machinery in the Red Alga Cyanidioschyzon merolae

Keiji Nishida, Osami Misumi, Fumi Yagisawa, Haruko Kuroiwa, Toshiyuki Nagata and Tsuneyoshi Kuroiwa

Department of Life Science, College of Science, Rikkyo University (IKN,OM,FY,HK,TK), and Department of Biological Sciences, Graduate School of Science, University of Tokyo (KN,FY,TN), Tokyo, Japan

Correspondence to: Keiji Nishida, Dept. of Life Science, College of Science, Rikkyo University, Tokyo 171-8501, Toshima-ku, Japan. E-mail: keiji{at}platz.jp

In the mitochondria of primitive eukaryotes, FtsZ and dynamin are part of the machinery involved in division of the inner and outer membranes, respectively. These genes also commonly function in the same manner during chloroplast division. In this study, a relationship between the localization of the inner and outer division machinery was directly shown for the first time. Triple immunofluorescent labeling was performed in the red alga Cyanidioschyzon merolae by a device using narrow bandpass filter sets and bright photostable dyes. FtsZ (CmFtsZ1) and dynamin (CmDnm1) localizations were examined simultaneously throughout the mitochondrial division cycle with an alternative mitochondrial marker protein, the mitochondrial translation elongation factor EF-Tu, whose localization was also shown to be identical to the mitochondrial matrix. FtsZ and dynamin did not necessarily co-localize when both were recruited to the mitochondrial constriction site, indicating that inner and outer dividing machineries are not in tight association during the late stage of division. (J Histochem Cytochem 52:843–849, 2004)

Key Words: FtsZ • dynamin • elongation factor • Tu • mitochondrial division • Cyanidioschyzon merolae • Alexa • triple immunofluorescence

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