Home LiteratureArticle Details
PMID: 15766329 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Silencing of p21-activated kinase attenuates vimentin phosphorylation on Ser-56 and reorientation of the vimentin network during stimulation of smooth muscle cells by 5-hydroxytryptamine.

The Biochemical journal ·Vol. 388 ·No. Pt 3 ·2005-06-15 ·Pages 773-83

Tang DD, Bai Y, Gunst SJ

Abstract

Vimentin intermediate filaments undergo spatial reorganization in endothelial cells and fibroblasts in response to stimulation with platelet-derived growth factor and epidermal growth factor. In the present study, the vimentin network exhibited a curved filamentous structure in unstimulated smooth muscle cells. Vimentin filaments became straight and were arranged along the long axis of cells upon stimulation with 5-hydroxytryptamine (5-HT; serotonin). Stimulation of smooth muscle cells with 5-HT also induced phosphorylation of vimentin on Ser-56. Treatment of cells with small interfering RNA selectively down-regulated the expression of PAK1 (p21-activated kinase 1) without affecting the content of smooth muscle alpha-actin. The silencing of PAK1 inhibited the site-specific phosphorylation and spatial rearrangement of the vimentin network in response to stimulation with 5-HT. Neither the disruption of stress fibres by cytochalasin D nor the inhibition of protein tyrosine phosphorylation affects the spatial reorganization of vimentin intermediate filaments in response to stimulation with 5-HT. In addition, stimulation of smooth muscle cells with 5-HT increased the ratio of soluble to insoluble vimentin. PAK1 silencing attenuated increases in the ratio of soluble to insoluble vimentin upon stimulation with 5-HT. These results suggest that the PAK-mediated site-specific phosphorylation of vimentin may play a role in regulating the reorganization of vimentin intermediate filaments during stimulation of smooth muscle cells with 5-HT.

MeSH Terms
Actin Cytoskeleton/chemistry,metabolism Animals Cells, Cultured Dogs Down-Regulation Gene Expression Regulation, Enzymologic Intermediate Filaments/chemistry,drug effects,metabolism Muscle, Smooth/cytology,drug effects,metabolism Phosphorylation/drug effects Phosphoserine/metabolism Protein Serine-Threonine Kinases/biosynthesis,deficiency,genetics,metabolism RNA Interference RNA, Small Interfering/genetics,metabolism Serotonin/pharmacology Solubility Trachea Vimentin/chemistry,metabolism p21-Activated Kinases
Chemicals
RNA, Small Interfering Vimentin Phosphoserine Serotonin Protein Serine-Threonine Kinases p21-Activated Kinases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Tang Dale D
Center for Cardiovascular Sciences, Albany Medical College, 47 New Scotland Avenue, MC-8, ME-424, Albany, NY 12208, USA. [email protected]
Bai Ying
Gunst Susan J
References (49)
49 references, click to expand
  1. Impaired flow-induced dilation in mesenteric resistance arteries from mice lacking vimentin.
    J Clin Invest. 1997 Dec 1;100(11):2909-14 PMID: 9389758
  2. Phosphorylation of vimentin by Rho-associated kinase at a unique amino-terminal site that is specifically phosphorylated during cytokinesis.
    J Biol Chem. 1998 May 8;273(19):11728-36 PMID: 9565595
  3. Tyrosine phosphorylation and association of p130Cas and c-Crk II by ANG II in vascular smooth muscle cells.
    Am J Physiol. 1998 Apr;274(4 Pt 2):H1059-65 PMID: 9575907
  4. Impaired mechanical stability, migration and contractile capacity in vimentin-deficient fibroblasts.
    J Cell Sci. 1998 Jul;111 ( Pt 13):1897-907 PMID: 9625752
  5. PDGF induces reorganization of vimentin filaments.
    J Cell Sci. 1998 Jul 30;111 ( Pt 14):1973-80 PMID: 9645945
  6. Mechanosensitive tyrosine phosphorylation of paxillin and focal adhesion kinase in tracheal smooth muscle.
    Am J Physiol. 1999 Jan;276(1 Pt 1):C250-8 PMID: 9886941
  7. Integrating the actin and vimentin cytoskeletons. adhesion-dependent formation of fimbrin-vimentin complexes in macrophages.
    J Cell Biol. 1999 Aug 23;146(4):831-42 PMID: 10459017
  8. Actin reorganization in airway smooth muscle cells involves Gq and Gi-2 activation of Rho.
    Am J Physiol. 1999 Sep;277(3 Pt 1):L653-61 PMID: 10484474
  9. Activation of the Arp2/3 complex by N-WASp is required for actin polymerization and contraction in smooth muscle.
    Am J Physiol Cell Physiol. 2005 May;288(5):C1145-60 PMID: 15625304
  10. Intermediate filaments and their associates: multi-talented structural elements specifying cytoarchitecture and cytodynamics.
    Curr Opin Cell Biol. 2000 Feb;12(1):79-90 PMID: 10679360
  11. Vimentin in cultured chromaffin cells: an immunofluorescent, biochemical and functional study.
    Cell Physiol Biochem. 2000;10(1-2):91-8 PMID: 10844400
  12. Contribution of intermediate filaments to cell stiffness, stiffening, and growth.
    Am J Physiol Cell Physiol. 2000 Jul;279(1):C188-94 PMID: 10898730
  13. Role of Rho in Ca(2+)-insensitive contraction and paxillin tyrosine phosphorylation in smooth muscle.
    Am J Physiol Cell Physiol. 2000 Aug;279(2):C308-18 PMID: 10912996
  14. Molecular mechanisms controlling actin filament dynamics in nonmuscle cells.
    Annu Rev Biophys Biomol Struct. 2000;29:545-76 PMID: 10940259
  15. Cdc42Hs and Rac1 GTPases induce the collapse of the vimentin intermediate filament network.
    J Biol Chem. 2000 Oct 20;275(42):33046-52 PMID: 10900195
  16. Paxillin and focal adhesion signalling.
    Nat Cell Biol. 2000 Dec;2(12):E231-6 PMID: 11146675
  17. Depletion of focal adhesion kinase by antisense depresses contractile activation of smooth muscle.
    Am J Physiol Cell Physiol. 2001 Apr;280(4):C874-83 PMID: 11245605
  18. Invited review: focal adhesion and small heat shock proteins in the regulation of actin remodeling and contractility in smooth muscle.
    J Appl Physiol (1985). 2001 Aug;91(2):963-72 PMID: 11457815
  19. Intercellular mechanotransduction: cellular circuits that coordinate tissue responses to mechanical loading.
    Biochem Biophys Res Commun. 2001 Aug 3;285(5):1077-83 PMID: 11478763
  20. Selected contribution: roles of focal adhesion kinase and paxillin in the mechanosensitive regulation of myosin phosphorylation in smooth muscle.
    J Appl Physiol (1985). 2001 Sep;91(3):1452-9 PMID: 11509548
  21. Exchange of the actin-bound nucleotide in intact arterial smooth muscle.
    J Biol Chem. 2001 Dec 21;276(51):48398-403 PMID: 11602582
  22. Pressure-induced actin polymerization in vascular smooth muscle as a mechanism underlying myogenic behavior.
    FASEB J. 2002 Jan;16(1):72-6 PMID: 11772938
  23. Stable suppression of gene expression by RNAi in mammalian cells.
    Proc Natl Acad Sci U S A. 2002 Feb 5;99(3):1443-8 PMID: 11818553
  24. Phosphorylation and reorganization of vimentin by p21-activated kinase (PAK).
    Genes Cells. 2002 Feb;7(2):91-7 PMID: 11895474
  25. p21-activated kinases: three more join the Pak.
    Int J Biochem Cell Biol. 2002 Jul;34(7):713-7 PMID: 11950587
  26. The focal adhesion protein paxillin regulates contraction in canine tracheal smooth muscle.
    J Physiol. 2002 Jul 15;542(Pt 2):501-13 PMID: 12122148
  27. Stiffness changes in cultured airway smooth muscle cells.
    Am J Physiol Cell Physiol. 2002 Sep;283(3):C792-801 PMID: 12176736
  28. Cdc42/Rac1-dependent activation of the p21-activated kinase (PAK) regulates human platelet lamellipodia spreading: implication of the cortical-actin binding protein cortactin.
    Blood. 2002 Dec 15;100(13):4462-9 PMID: 12453877
  29. Vimentin intermediate filament reorganization by Cdc42: involvement of PAK and p70 S6 kinase.
    Eur J Cell Biol. 2002 Dec;81(12):692-701 PMID: 12553669
  30. Calcium-independent contraction and sensitization of airway smooth muscle by p21-activated protein kinase.
    Am J Physiol Lung Cell Mol Physiol. 2003 May;284(5):L863-70 PMID: 12513968
  31. Downregulation of profilin with antisense oligodeoxynucleotides inhibits force development during stimulation of smooth muscle.
    Am J Physiol Heart Circ Physiol. 2003 Oct;285(4):H1528-36 PMID: 12805028
  32. Role of Crk-associated substrate in the regulation of vascular smooth muscle contraction.
    Hypertension. 2003 Oct;42(4):858-63 PMID: 12885796
  33. Asymmetry in the assembly of the RNAi enzyme complex.
    Cell. 2003 Oct 17;115(2):199-208 PMID: 14567917
  34. Expression of non-phosphorylatable paxillin mutants in canine tracheal smooth muscle inhibits tension development.
    J Physiol. 2003 Nov 15;553(Pt 1):21-35 PMID: 12949231
  35. Tension development during contractile stimulation of smooth muscle requires recruitment of paxillin and vinculin to the membrane.
    Am J Physiol Cell Physiol. 2004 Feb;286(2):C433-47 PMID: 14576084
  36. Specific in vivo phosphorylation sites determine the assembly dynamics of vimentin intermediate filaments.
    J Cell Sci. 2004 Feb 29;117(Pt 6):919-32 PMID: 14762106
  37. Novel p21-activated kinase-dependent protrusions characteristically formed at the edge of transformed cells.
    Exp Cell Res. 2004 May 1;295(2):432-47 PMID: 15093742
  38. The contractile apparatus of vascular smooth muscle: intermediate high voltage stereo electron microscopy.
    J Mol Biol. 1975 Oct 15;98(1):17-29 PMID: 1195377
  39. Identification of a distinct soluble subunit of an intermediate filament protein: tetrameric vimentin from living cells.
    Proc Natl Acad Sci U S A. 1985 Dec;82(23):7929-33 PMID: 3865206
  40. Angiotensin II stimulates vimentin phosphorylation via a Ca2+-dependent, protein kinase C-independent mechanism in cultured vascular smooth muscle cells.
    J Biol Chem. 1988 Dec 25;263(36):19758-63 PMID: 3143730
  41. Comparison of the mitogenic activity of angiotensin II and serotonin on porcine arterial smooth muscle cells.
    Atherosclerosis. 1991 Jun;88(2-3):213-8 PMID: 1892486
  42. A brain serine/threonine protein kinase activated by Cdc42 and Rac1.
    Nature. 1994 Jan 6;367(6458):40-6 PMID: 8107774
  43. Visualization and function of vimentin phosphorylation by cdc2 kinase during mitosis.
    J Biol Chem. 1994 Dec 9;269(49):31097-106 PMID: 7983050
  44. Activation of the small GTP-binding proteins rho and rac by growth factor receptors.
    J Cell Sci. 1995 Jan;108 ( Pt 1):225-33 PMID: 7738099
  45. Structure-function relations of smooth muscle calponin. The critical role of serine 175.
    J Biol Chem. 1996 Apr 12;271(15):8605-11 PMID: 8621490
  46. Structure-function relationships in smooth muscle: the missing links.
    Bioessays. 1995 Sep;17(9):785-92 PMID: 8763831
  47. Markers of airway smooth muscle cell phenotype.
    Am J Physiol. 1996 Jun;270(6 Pt 1):L1040-51 PMID: 8764231
  48. Intermediate filament proteins in adult human arteries.
    Anat Rec. 1997 Apr;247(4):439-48 PMID: 9096782
  49. Rho, Rac and Cdc42 regulate actin organization and cell adhesion in macrophages.
    J Cell Sci. 1997 Mar;110 ( Pt 6):707-20 PMID: 9099945
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
1470-8728
DOI
10.1042/BJ20050065
Email
[email protected]
Published
2005-06-15
Pages
773-83
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1183456
Subset
IM
Grants
NHLBI NIH HHS · R01 HL029289-19 · United States
NHLBI NIH HHS · R01 HL075388 · United States
NHLBI NIH HHS · R01 HL074099 · United States
NHLBI NIH HHS · R01 HL029289 · United States
NHLBI NIH HHS · R01 HL074099-03 · United States
NHLBI NIH HHS · HL-75388 · United States
NHLBI NIH HHS · HL-29289 · United States
NHLBI NIH HHS · HL-074099 · United States
External Links
PubMed source DOI: 10.1042/BJ20050065 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2