Paxillin Regulates Rab5-mediated Vesicle Motility Through Modulating Microtubule Acetylation

Overview

Journal Mol Biol Cell

Specialties Cell Biology
Molecular Biology

Date 2023 Apr 12

PMID 37043310

Authors

Nicholas M Zehrbach

Fatemeh Dubois

Christopher E Turner

Affiliations

Soon will be listed here.

Abstract

Rab GTPase-mediated vesicle trafficking of cell surface proteins, including integrins, through endocytic and recycling pathways is important in controlling cell-extracellular matrix interactions during cell migration. The focal adhesion adaptor protein, paxillin, plays a central role in regulating adhesion dynamics and was previously shown to promote anterograde vesicle trafficking through modulation of microtubule acetylation via its inhibition of the deacetylase HDAC6. The role of paxillin in retrograde trafficking is unknown. Herein, we identified a role for paxillin in the modulation of the Rab5 GTPase, which is necessary for regulating early endosome dynamics and focal adhesion turnover. Using MDA-MB-231 breast cancer cells and paxillin (-/-) fibroblasts, paxillin was shown to impact Rab5-associated vesicle size and distribution, as well as Rab5 GTPase activity, through its modulation of HDAC6. Using a combination of real-time imaging and particle tracking analysis, paxillin was shown to promote Rab5-associated vesicle motility through inhibition of HDAC6-mediated micro-tubule deacetylation, along with the localization of active integrin to focal adhesions.

Citing Articles

A comparative analysis of paxillin and Hic-5 proximity interactomes.

Brock K, Alpha K, Brennan G, De Jong E, Luke E, Turner C Cytoskeleton (Hoboken). 2024; 82(1-2):12-31.

PMID: 38801098 PMC: 11599474. DOI: 10.1002/cm.21878.

References

Matteoni R, Kreis T . Translocation and clustering of endosomes and lysosomes depends on microtubules. J Cell Biol. 1987; 105(3):1253-65. PMC: 2114818. DOI: 10.1083/jcb.105.3.1253. View

Deakin N, Turner C . Distinct roles for paxillin and Hic-5 in regulating breast cancer cell morphology, invasion, and metastasis. Mol Biol Cell. 2010; 22(3):327-41. PMC: 3031464. DOI: 10.1091/mbc.E10-09-0790. View

Huang J, Imamura T, Olefsky J . Insulin can regulate GLUT4 internalization by signaling to Rab5 and the motor protein dynein. Proc Natl Acad Sci U S A. 2001; 98(23):13084-9. PMC: 60828. DOI: 10.1073/pnas.241368698. View

De Franceschi N, Hamidi H, Alanko J, Sahgal P, Ivaska J . Integrin traffic - the update. J Cell Sci. 2015; 128(5):839-52. PMC: 4342575. DOI: 10.1242/jcs.161653. View

Even A, Morelli G, Broix L, Scaramuzzino C, Turchetto S, Gladwyn-Ng I . ATAT1-enriched vesicles promote microtubule acetylation via axonal transport. Sci Adv. 2020; 5(12):eaax2705. PMC: 6920029. DOI: 10.1126/sciadv.aax2705. View

Xu W, Gulvady A, Goreczny G, Olson E, Turner C . Paxillin-dependent regulation of apical-basal polarity in mammary gland morphogenesis. Development. 2019; 146(9). PMC: 6526716. DOI: 10.1242/dev.174367. View

Dong J, Tay F, Swa H, Gunaratne J, Leung T, Burke B . Proximity biotinylation provides insight into the molecular composition of focal adhesions at the nanometer scale. Sci Signal. 2016; 9(432):rs4. DOI: 10.1126/scisignal.aaf3572. View

Brown M, Cary L, Jamieson J, Cooper J, Turner C . Src and FAK kinases cooperate to phosphorylate paxillin kinase linker, stimulate its focal adhesion localization, and regulate cell spreading and protrusiveness. Mol Biol Cell. 2005; 16(9):4316-28. PMC: 1196340. DOI: 10.1091/mbc.e05-02-0131. View

Tinevez J, Perry N, Schindelin J, Hoopes G, Reynolds G, Laplantine E . TrackMate: An open and extensible platform for single-particle tracking. Methods. 2016; 115:80-90. DOI: 10.1016/j.ymeth.2016.09.016. View

10.

Lawe D, Patki V, Lambright D, Corvera S . The FYVE domain of early endosome antigen 1 is required for both phosphatidylinositol 3-phosphate and Rab5 binding. Critical role of this dual interaction for endosomal localization. J Biol Chem. 2000; 275(5):3699-705. DOI: 10.1074/jbc.275.5.3699. View

11.

Bance B, Seetharaman S, Leduc C, Boeda B, Etienne-Manneville S . Microtubule acetylation but not detyrosination promotes focal adhesion dynamics and astrocyte migration. J Cell Sci. 2019; 132(7). DOI: 10.1242/jcs.225805. View

12.

Barbieri M, Li G, Mayorga L, Stahl P . Characterization of Rab5:Q79L-stimulated endosome fusion. Arch Biochem Biophys. 1996; 326(1):64-72. DOI: 10.1006/abbi.1996.0047. View

13.

Brown M, Perrotta J, Turner C . Identification of LIM3 as the principal determinant of paxillin focal adhesion localization and characterization of a novel motif on paxillin directing vinculin and focal adhesion kinase binding. J Cell Biol. 1996; 135(4):1109-23. PMC: 2133378. DOI: 10.1083/jcb.135.4.1109. View

14.

Palamidessi A, Frittoli E, Ducano N, Offenhauser N, Sigismund S, Kajiho H . The GTPase-activating protein RN-tre controls focal adhesion turnover and cell migration. Curr Biol. 2013; 23(23):2355-64. DOI: 10.1016/j.cub.2013.09.060. View

15.

Webster D, Borisy G . Microtubules are acetylated in domains that turn over slowly. J Cell Sci. 1989; 92 ( Pt 1):57-65. DOI: 10.1242/jcs.92.1.57. View

16.

Turner C . Paxillin and focal adhesion signalling. Nat Cell Biol. 2001; 2(12):E231-6. DOI: 10.1038/35046659. View

17.

Palazzo A, Eng C, Schlaepfer D, Marcantonio E, Gundersen G . Localized stabilization of microtubules by integrin- and FAK-facilitated Rho signaling. Science. 2004; 303(5659):836-9. DOI: 10.1126/science.1091325. View

18.

Soppina V, Rai A, Ramaiya A, Barak P, Mallik R . Tug-of-war between dissimilar teams of microtubule motors regulates transport and fission of endosomes. Proc Natl Acad Sci U S A. 2009; 106(46):19381-6. PMC: 2770008. DOI: 10.1073/pnas.0906524106. View

19.

Hendricks A, Perlson E, Ross J, Schroeder 3rd H, Tokito M, Holzbaur E . Motor coordination via a tug-of-war mechanism drives bidirectional vesicle transport. Curr Biol. 2010; 20(8):697-702. PMC: 2908734. DOI: 10.1016/j.cub.2010.02.058. View

20.

Janke C, Montagnac G . Causes and Consequences of Microtubule Acetylation. Curr Biol. 2017; 27(23):R1287-R1292. DOI: 10.1016/j.cub.2017.10.044. View