A Raft-derived, Pak1-regulated Entry Participates in Alpha2beta1 Integrin-dependent Sorting to Caveosomes

Overview

Journal Mol Biol Cell

Specialties Cell Biology
Molecular Biology

Date 2008 May 2

PMID 18448666

Citations 54

Authors

Mikko Karjalainen

Elina Kakkonen

Paula Upla

Heli Paloranta

Pasi Kankaanpaa

Prisca Liberali

G Herma Renkema

Timo Hyypia

Jyrki Heino

Varpu Marjomaki

Affiliations

Soon will be listed here.

Abstract

We have previously shown that a human picornavirus echovirus 1 (EV1) is transported to caveosomes during 2 h together with its receptor alpha2beta1 integrin. Here, we show that the majority of early uptake does not occur through caveolae. alpha2beta1 integrin, clustered by antibodies or by EV1 binding, is initially internalized from lipid rafts into tubulovesicular structures. These vesicles accumulate fluid-phase markers but do not initially colocalize with caveolin-1 or internalized simian virus 40 (SV40). Furthermore, the internalized endosomes do not contain glycosylphosphatidylinositol (GPI)-anchored proteins or flotillin 1, suggesting that clustered alpha2beta1 integrin does not enter the GPI-anchored protein enriched endosomal compartment or flotillin pathways, respectively. Endosomes mature further into larger multivesicular bodies between 15 min to 2 h and concomitantly recruit caveolin-1 or SV40 inside. Cell entry is regulated by p21-activated kinase (Pak)1, Rac1, phosphatidylinositol 3-kinase, phospholipase C, and actin but not by dynamin 2 in SAOS-alpha2beta1 cells. An amiloride analog, 5-(N-ethyl-N-isopropanyl) amiloride, blocks infection, causes integrin accumulation in early tubulovesicular structures, and prevents their structural maturation into multivesicular structures. Our results together suggest that alpha2beta1 integrin clustering defines its own entry pathway that is Pak1 dependent but clathrin and caveolin independent and that is able to sort cargo to caveosomes.

Citing Articles

Covalent and non-covalent coupling of a Au nanocluster with a fluorophore: energy transfer, quenching and intracellular pH sensing.

Hulkko E, Lahtinen T, Marjomaki V, Pohjolainen E, Saarnio V, Sokolowska K Nanoscale Adv. 2022; 3(23):6649-6658.

PMID: 36132657 PMC: 9417352. DOI: 10.1039/d1na00368b.

Multi-Omics Analysis of Glioblastoma Cells' Sensitivity to Oncolytic Viruses.

Lipatova A, Soboleva A, Gorshkov V, Bubis J, Solovyeva E, Krasnov G Cancers (Basel). 2021; 13(21).

PMID: 34771433 PMC: 8582528. DOI: 10.3390/cancers13215268.

6-Thioguanine Inhibits Herpes Simplex Virus 1 Infection of Eyes.

Chen D, Liu Y, Zhang F, You Q, Ma W, Wu J Microbiol Spectr. 2021; 9(3):e0064621.

PMID: 34730435 PMC: 8567252. DOI: 10.1128/Spectrum.00646-21.

Establishment of an Endocytosis-Related Prognostic Signature for Patients With Low-Grade Glioma.

Wang D, Liu S, Wang G Front Genet. 2021; 12:709666.

PMID: 34552618 PMC: 8450508. DOI: 10.3389/fgene.2021.709666.

Rac1-PAK1 regulation of Rab11 cycling promotes junction destabilization.

Erasmus J, Smolarczyk K, Brezovjakova H, Mohd-Naim N, Lozano E, Matter K J Cell Biol. 2021; 220(6).

PMID: 33914026 PMC: 8091128. DOI: 10.1083/jcb.202002114.

References

Liberali P, Kakkonen E, Turacchio G, Valente C, Spaar A, Perinetti G . The closure of Pak1-dependent macropinosomes requires the phosphorylation of CtBP1/BARS. EMBO J. 2008; 27(7):970-81. PMC: 2323256. DOI: 10.1038/emboj.2008.59. View

Glebov O, Bright N, Nichols B . Flotillin-1 defines a clathrin-independent endocytic pathway in mammalian cells. Nat Cell Biol. 2005; 8(1):46-54. DOI: 10.1038/ncb1342. View

Amyere M, Payrastre B, Krause U, Van Der Smissen P, Veithen A, Courtoy P . Constitutive macropinocytosis in oncogene-transformed fibroblasts depends on sequential permanent activation of phosphoinositide 3-kinase and phospholipase C. Mol Biol Cell. 2000; 11(10):3453-67. PMC: 15006. DOI: 10.1091/mbc.11.10.3453. View

Wadia J, Stan R, Dowdy S . Transducible TAT-HA fusogenic peptide enhances escape of TAT-fusion proteins after lipid raft macropinocytosis. Nat Med. 2004; 10(3):310-5. DOI: 10.1038/nm996. View

Manser E, Huang H, Loo T, Chen X, Dong J, Leung T . Expression of constitutively active alpha-PAK reveals effects of the kinase on actin and focal complexes. Mol Cell Biol. 1997; 17(3):1129-43. PMC: 231838. DOI: 10.1128/MCB.17.3.1129. View

Sabharanjak S, Sharma P, Parton R, Mayor S . GPI-anchored proteins are delivered to recycling endosomes via a distinct cdc42-regulated, clathrin-independent pinocytic pathway. Dev Cell. 2002; 2(4):411-23. DOI: 10.1016/s1534-5807(02)00145-4. View

Papakonstanti E, Stournaras C . Association of PI-3 kinase with PAK1 leads to actin phosphorylation and cytoskeletal reorganization. Mol Biol Cell. 2002; 13(8):2946-62. PMC: 117954. DOI: 10.1091/mbc.02-01-0599. View

Bonazzi M, Spano S, Turacchio G, Cericola C, Valente C, Colanzi A . CtBP3/BARS drives membrane fission in dynamin-independent transport pathways. Nat Cell Biol. 2005; 7(6):570-80. DOI: 10.1038/ncb1260. View

Pietiainen V, Marjomaki V, Upla P, Pelkmans L, Helenius A, Hyypia T . Echovirus 1 endocytosis into caveosomes requires lipid rafts, dynamin II, and signaling events. Mol Biol Cell. 2004; 15(11):4911-25. PMC: 524743. DOI: 10.1091/mbc.e04-01-0070. View

10.

Nakase I, Niwa M, Takeuchi T, Sonomura K, Kawabata N, Koike Y . Cellular uptake of arginine-rich peptides: roles for macropinocytosis and actin rearrangement. Mol Ther. 2004; 10(6):1011-22. DOI: 10.1016/j.ymthe.2004.08.010. View

11.

Marjomaki V, Pietiainen V, Matilainen H, Upla P, Ivaska J, Nissinen L . Internalization of echovirus 1 in caveolae. J Virol. 2002; 76(4):1856-65. PMC: 135881. DOI: 10.1128/jvi.76.4.1856-1865.2002. View

12.

Mettlen M, Platek A, Van Der Smissen P, Carpentier S, Amyere M, Lanzetti L . Src triggers circular ruffling and macropinocytosis at the apical surface of polarized MDCK cells. Traffic. 2006; 7(5):589-603. DOI: 10.1111/j.1600-0854.2006.00412.x. View

13.

Xing L, Huhtala M, Pietiainen V, Kapyla J, Vuorinen K, Marjomaki V . Structural and functional analysis of integrin alpha2I domain interaction with echovirus 1. J Biol Chem. 2004; 279(12):11632-8. DOI: 10.1074/jbc.M312441200. View

14.

Mayor S, Pagano R . Pathways of clathrin-independent endocytosis. Nat Rev Mol Cell Biol. 2007; 8(8):603-12. PMC: 7617177. DOI: 10.1038/nrm2216. View

15.

Fivaz M, Vilbois F, Thurnheer S, Pasquali C, Abrami L, Bickel P . Differential sorting and fate of endocytosed GPI-anchored proteins. EMBO J. 2002; 21(15):3989-4000. PMC: 126144. DOI: 10.1093/emboj/cdf398. View

16.

Kalia M, Kumari S, Chadda R, Hill M, Parton R, Mayor S . Arf6-independent GPI-anchored protein-enriched early endosomal compartments fuse with sorting endosomes via a Rab5/phosphatidylinositol-3'-kinase-dependent machinery. Mol Biol Cell. 2006; 17(8):3689-704. PMC: 1525230. DOI: 10.1091/mbc.e05-10-0980. View

17.

Ivaska J, Reunanen H, Westermarck J, Koivisto L, Kahari V, Heino J . Integrin alpha2beta1 mediates isoform-specific activation of p38 and upregulation of collagen gene transcription by a mechanism involving the alpha2 cytoplasmic tail. J Cell Biol. 1999; 147(2):401-16. PMC: 2174225. DOI: 10.1083/jcb.147.2.401. View

18.

Upla P, Marjomaki V, Kankaanpaa P, Ivaska J, Hyypia T, van der Goot F . Clustering induces a lateral redistribution of alpha 2 beta 1 integrin from membrane rafts to caveolae and subsequent protein kinase C-dependent internalization. Mol Biol Cell. 2003; 15(2):625-36. PMC: 329276. DOI: 10.1091/mbc.e03-08-0588. View

19.

White I, Bailey L, Aghakhani M, Moss S, Futter C . EGF stimulates annexin 1-dependent inward vesiculation in a multivesicular endosome subpopulation. EMBO J. 2005; 25(1):1-12. PMC: 1356341. DOI: 10.1038/sj.emboj.7600759. View

20.

Marjomaki V, Huovila A, Surkka M, Jokinen I, Salminen A . Lysosomal trafficking in rat cardiac myocytes. J Histochem Cytochem. 1990; 38(8):1155-64. DOI: 10.1177/38.8.2164059. View