FAM123A Binds to Microtubules and Inhibits the Guanine Nucleotide Exchange Factor ARHGEF2 to Decrease Actomyosin Contractility

Priscila F. Siesser, Marta Motolese, Matthew P. Walker, Dennis Goldfarb, Kelly Gewain, Feng Yan, Rima M. Kulikauskas, Andy J. Chien, Linda Wordeman, Michael B. Major*

*To whom correspondence should be addressed. E-mail: benmajor{at}med.unc.edu

This PDF file includes:

• Fig. S1. FAM123C does not localize to the cytoplasmic membrane.
• Fig. S2. FAM123A binds FBXW11.
• Fig. S3. Mutation of the SKIP motif in FAM123A abrogates its microtubule localization and tracking.
• Fig. S4. FAM123A overexpression alters the subcellular localization of EB1.
• Fig. S5. EB1 and microtubule association is dispensable for the FAM123A control of β-catenin–dependent WNT signaling.
• Fig. S6. GEF-H1 silencing alters microtubule organization and actomyosin contractility.
• Fig. S7. The increased actin contractility observed in FAM123A siRNA–transfected cells is due to the specific silencing of FAM123A.
• Fig. S8. FAM123A controls maturation of focal adhesions during cell spreading.
• Legends for Movies S1 to S7

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Other Supplementary Material for this manuscript includes the following:

• Table S1 (Microsoft Excel format). Affinity purification and MS data.
• Movie S1 (.mov format). Venus-fused FAM123A localizes to the cell membrane and to filamentous structures in the cell lamellipodium.
• Movie S2 (.mov format). Venus-fused WTX localizes predominantly to the cell membrane.
• Movie S3 (.mov format). EGFP-fused FAM123A moves on presumed microtubules toward the cell cortex.
• Movie S4 (.mov format). EGFP-fused FAM123A tracks the tips and distal ends of EB3-associated microtubules.
• Movie S5 (.mov format). FAM123A-IPAA shows decreased EB3-associated plus-end tip tracking and decoration of distal ends of microtubules.
• Movie S6 (.mov format). The C-terminal fragment of FAM123A, which contains the SKIP EB-interaction motif, behaves as a classic +TIP showing robust microtubule tip tracking, but does not decorate the microtubule lattice.
• Movie S7 (.mov format). The C-terminal fragment of FAM123A with a mutated SKIP motif does not show colocalization or tip tracking of microtubules.

© 2012 American Association for the Advancement of Science