Interaction of PDZRhoGEF with microtubule-associated protein 1 light chains: Link between microtubules, actin cytoskeleton, and neuronal polarity

David M. Longhurst, Mitsunori Watanabe, Jeffrey D. Rothstein, Mandy Jackson

Research output: Contribution to journalArticlepeer-review

21 Scopus citations


Rat (r) PDZRhoGEF, initially identified as a glutamate transporter EAAT4-associated protein, is amember of a novel RhoGEF subfamily. The N terminus of the protein contains a PDZ and a proline-rich domain, two motifs known to be involved in protein-protein interactions. By using the yeast two-hybrid approach, we screened for proteins that interact with the N terminus of rPDZRhoGEF. The light chain 2 of microtubule-associated protein 1 (LC2) was the only protein identified from the screen that does not contain a type I PDZ-binding motif at its extreme C terminus (-(S/T)Xφ-COOH, where φ is a hydrophobic amino acid). However, the C terminus does conform to a type II-binding motif (-φXφ). We report here that rPDZRhoGEF interacts with LC2 via the PDZ domain, and the interaction is abolished by mutations in the carboxylate-binding loop. The specificity of the interaction was confirmed using GST fusion protein pull-down assays and coimmunoprecipitations. Expression of rPDZRhoGEF mutants that are unable to interact with proteins via the carboxylate-binding loop induced changes in cell morphology and actin organization. These mutants alter the activation of RhoGTPases, and coexpression of dominant-negative RhoGTPases prevent the morphological changes. Furthermore, in cells expressing wild type rPDZRhoGEF, drug-induced microtubule depolymerization produces changes in cell morphology that are similar to those induced by rPDZRhoGEF mutants. These results indicate that modulation of the guanine nucleotide exchange activity of rPDZRhoGEF through interaction with microtubule-associated protein light chains may coordinate microtubule integrity and the reorganization of actin cytoskeleton. This coordinated action of the actin and microtubular cytoskeletons is essential for the development and maintenance of neuronal polarity.

Original languageEnglish (US)
Pages (from-to)12030-12040
Number of pages11
JournalJournal of Biological Chemistry
Issue number17
StatePublished - Apr 28 2006

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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