Characterization of the arabidopsis formin-like protein AFH1 and its interacting protein.
A partial cDNA encoding an Arabidopsis thaliana FH (Formin Homology) protein (AFH1) was used as a probe to clone a full length AFH1 cDNA. The deduced protein encoded by the cDNA contains a FH1 domain rich in proline residues and a C-terminal FH2 domain which is highly conserved amongst FH proteins. In contrast to FH proteins of other organisms, the predicted AFH1 protein also contains a putative signal peptide and a transmembrane domain suggesting its association with membrane. Cell fractionation by differential centrifugation demonstrated the presence of AFH1 in the Triton X-100 insoluble microsomal fraction. An Arabidopsis cDNA library was screened to identify proteins that interact with the C-terminal region of AFH1 using yeast two-hybrid assays, and one of the isolated cDNAs encoded a novel protein, FIP2. Experiments using recombinant proteins expressed in E. coli demonstrated that FIP2 interacted directly with AFH1. The amino acid sequence of FIP2 has partial homology to bacterial putative membrane proteins and animal A-type K+ ATPases. AFH1 may form a membrane anchored complex with FIP2, which might be involved in the organization of the actin cytoskeleton.
Laboratory of Plant Molecular Biology, The Rockefeller University, New York, NY 10021, USA.
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D000251:Adenosine Triphosphatases / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins; D002352:Carrier Proteins / Q000378:metabolism*; D027682:Cation Transport Proteins; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary; D004926:Escherichia coli; D000080865:Formins; D015723:Gene Library; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008861:Microsomes / Q000378:metabolism; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
C417969:AFH1 protein, Arabidopsis; D029681:Arabidopsis Proteins; D002352:Carrier Proteins; D027682:Cation Transport Proteins; D018076:DNA, Complementary; C417970:FIP2 protein, Arabidopsis; D000080865:Formins; D008565:Membrane Proteins; D010940:Plant Proteins; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D000251:Adenosine Triphosphatases; C022761:potassium transporting ATPase