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Eukaryotic Cell, July 2005, p. 1253-1263, Vol. 4, No. 7
1535-9778/05/$08.00+0     doi:10.1128/EC.4.7.1253-1263.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Centrin Scaffold in Chlamydomonas reinhardtii Revealed by Immunoelectron Microscopy

Stefan Geimer^* and Michael Melkonian

Botanisches Institut, Universität zu Köln, D-50931 Köln, Germany

Received 7 March 2005/ Accepted 10 May 2005

In the flagellate green alga Chlamydomonas reinhardtii the Ca²⁺-binding EF-hand protein centrin is encoded by a single-copy gene. Previous studies have localized the protein to four distinct structures in the flagellar apparatus: the nucleus-basal body connector, the distal connecting fiber, the flagellar transitional region, and the axoneme. To explain the disjunctive distribution of centrin, the interaction of centrin with as yet unknown specific centrin-binding proteins has been implied. Here, we demonstrate using serial section postembedding immunoelectron microscopy of isolated cytoskeletons that centrin is located in additional structures (transitional fibers and basal body lumen) and that the centrin-containing structures of the basal apparatus are likely part of a continuous filamentous scaffold that extends from the nucleus to the flagellar bases. In addition, we show that centrin is located in the distal lumen of the basal body in a rotationally asymmetric structure, the V-shaped filament system. This novel centrin-containing structure has also been detected near the distal end of the probasal bodies. Taken together, these results suggest a role for a rotationally asymmetric centrin "seed" in the growth and development of the centrin scaffold following replication of the basal apparatus.

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* Corresponding author. Mailing address: Universität Bayreuth, Biologie/Elektronenmikroskopie NW I / B 1, Universitätsstr. 30, D-95447 Bayreuth, Germany. Phone: 49 921 55 2164. Fax: 49 921 55 4301. E-mail: stefan.geimer{at}uni-bayreuth.de.

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Eukaryotic Cell, July 2005, p. 1253-1263, Vol. 4, No. 7
1535-9778/05/$08.00+0     doi:10.1128/EC.4.7.1253-1263.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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