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Localization of two homologous Arabidopsis kinesin-related proteins in the phragmoplast

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Abstract

During plant cytokinesis, kinesin-related motor proteins are believed to play critical roles in microtubule organization and vesicle transport in the phragmoplast. Previously, we reported that the motor AtPAKRP1 was associated with the plus end of phragmoplast microtubules in Arabidopsis thaliana [Lee Y-RJ, Liu B (2000) Curr Biol 10:797–800]. In this paper, we report a full-length cDNA from the same organism, which encodes a polypeptide 74% identical to AtPAKRP1. This AtPAKRP1-like protein—AtPAKRP1L—and AtPAKRP1 share similar domain structures along the polypeptides. Peptide antibodies were raised and purified to distinguish the two polypeptides in vitro and in vivo. When monospecific anti-AtPAKRP1 and anti-AtPAKRP1L antibodies were used in immunofluorescence, they both decorated the plus end of phragmoplast microtubules at all stages of phragmoplast development. Their localization patterns were indistinguishable from each other. By using bacterially expressed fusion proteins of motor-less versions of both polypeptides, it was revealed that AtPAKRP1 and AtPAKRP1L were able to interact with themselves and with each other. Using T-DNA insertional mutants, it was also demonstrated that AtPAKRP1 and AtPAKRP1L were not required for each other’s localization. Our results therefore indicate that AtPAKRP1 and AtPAKRP1L are both expressed in the same cells, and likely have identical functions in the phragmoplast by forming either homodimers or heterodimers.

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Abbreviations

AtPAKRP1 :

Arabidopsis thaliana phragmoplast-associated kinesin-related protein 1

AtPAKRP1L :

A. thaliana phragmoplast-associated kinesin-related protein 1-like

GST :

Glutathione S-transferase

KRP :

Kinesin-related protein

6×His :

Six-histidine tag

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Acknowledgements

This work was supported by the United States Department of Agriculture (grant no. CSREES/NRICGP 2001-35304-01913). We would like to thank the Kazusa DNA Institute, Arabidopsis Biological Research Center, the University of Wisconsin Biotechnology Center, and the Syngenta Terrey Mesa Research Institute for providing cDNA clones and mutant screening services.

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  1. Section of Plant Biology, University of California, One Shields Avenue, Davis, CA 95616, USA

    Ruiqin Pan, Y.-R. Julie Lee & Bo Liu

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  1. Ruiqin Pan
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  2. Y.-R. Julie Lee
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  3. Bo Liu
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Correspondence to Bo Liu.

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Pan, R., Lee, YR.J. & Liu, B. Localization of two homologous Arabidopsis kinesin-related proteins in the phragmoplast. Planta 220, 156–164 (2004). https://doi.org/10.1007/s00425-004-1324-4

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