The EF-hand Ca(2+)-binding protein p22 associates with microtubules in an N-myristoylation-dependent manner.
Proteins containing the EF-hand Ca(2+)-binding motif, such as calmodulin and calcineurin B, function as regulators of various cellular processes. Here we focus on p22, an N-myristoylated, widely expressed EF-hand Ca(2+)-binding protein conserved throughout evolution, which was shown previously to be required for membrane traffic. Immunofluorescence studies show that p22 distributes along microtubules during interphase and mitosis in various cell lines. Moreover, we report that p22 associates with the microtubule cytoskeleton indirectly via a cytosolic microtubule-binding factor. Gel filtration studies indicate that the p22-microtubule-binding activity behaves as a 70- to 30-kDa globular protein. Our results indicate that p22 associates with microtubules via a novel N-myristoylation-dependent mechanism that does not involve classic microtubule-associated proteins and motor proteins. The association of p22 with microtubules requires the N-myristoylation of p22 but does not involve p22's Ca(2+)-binding activity, suggesting that the p22-microtubule association and the role of p22 in membrane traffic are functionally related, because N-myristoylation is required for both events. Therefore, p22 is an excellent candidate for a protein that can mediate interactions between the microtubule cytoskeleton and membrane traffic.
Timm
S
S
Department of Biology, University of Virginia, Charlottesville, Virginia 22903, USA.
Titus
B
B
Bernd
K
K
Barroso
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D002453:Cell Cycle; D002460:Cell Line; D003600:Cytosol / Q000378:metabolism; D020832:EF Hand Motifs; D005455:Fluorescent Antibody Technique; D008074:Lipoproteins / Q000235:genetics / Q000378:metabolism*; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D008970:Molecular Weight; D019814:Myristic Acid / Q000378:metabolism*; D010446:Peptide Fragments / Q000276:immunology; D011485:Protein Binding; D051381:Rats; D011994:Recombinant Proteins; D012965:Sodium Chloride / Q000494:pharmacology
D002135:Calcium-Binding Proteins; D008074:Lipoproteins; D008869:Microtubule-Associated Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; C098929:calcium binding protein p22, rat; D019814:Myristic Acid; D012965:Sodium Chloride