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PMID: 15699215 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Structural determinants for EB1-mediated recruitment of APC and spectraplakins to the microtubule plus end.

The Journal of cell biology ·Vol. 168 ·No. 4 ·2005-02-14 ·Pages 587-98

Slep KC, Rogers SL, Elliott SL, Ohkura H, Kolodziej PA, Vale RD

Abstract

EB1 is a member of a conserved protein family that localizes to growing microtubule plus ends. EB1 proteins also recruit cell polarity and signaling molecules to microtubule tips. However, the mechanism by which EB1 recognizes cargo is unknown. Here, we have defined a repeat sequence in adenomatous polyposis coli (APC) that binds to EB1's COOH-terminal domain and identified a similar sequence in members of the microtubule actin cross-linking factor (MACF) family of spectraplakins. We show that MACFs directly bind EB1 and exhibit EB1-dependent plus end tracking in vivo. To understand how EB1 recognizes APC and MACFs, we solved the crystal structure of the EB1 COOH-terminal domain. The structure reveals a novel homodimeric fold comprised of a coiled coil and four-helix bundle motif. Mutational analysis reveals that the cargo binding site for MACFs maps to a cluster of conserved residues at the junction between the coiled coil and four-helix bundle. These results provide a structural understanding of how EB1 binds two regulators of microtubule-based cell polarity.

MeSH Terms
Adenomatous Polyposis Coli Protein/genetics,metabolism Amino Acid Motifs/physiology Amino Acid Sequence Animals Binding Sites/physiology Cells, Cultured Drosophila Microfilament Proteins/metabolism Microtubule-Associated Proteins/metabolism Microtubules/metabolism Models, Molecular Molecular Sequence Data Mutagenesis/genetics Protein Binding
Chemicals
Adenomatous Polyposis Coli Protein EB1 microtubule binding proteins Microfilament Proteins Microtubule-Associated Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Slep Kevin C
The Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA 94107, USA.
Rogers Stephen L
Elliott Sarah L
Ohkura Hiroyuki
Kolodziej Peter A
Vale Ronald D
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200410114
Published
2005-02-14
Epub
2005-00-07
Pages
587-98
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2171753
Subset
IM
Grants
Wellcome Trust · United Kingdom
Wellcome Trust · 051091 · United Kingdom
NIGMS NIH HHS · R01 GM038499 · United States
PHS HHS · NIH38499 · United States
External Links
PubMed source DOI: 10.1083/jcb.200410114 PMC Full Text View journal details
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