Home LiteratureArticle Details
PMID: 9822607 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Targeted downregulation of caveolin-1 is sufficient to drive cell transformation and hyperactivate the p42/44 MAP kinase cascade.

The EMBO journal ·Vol. 17 ·No. 22 ·1998-11-16 ·Pages 6633-48

Galbiati F, Volonte D, Engelman JA, Watanabe G, Burk R, Pestell RG, Lisanti MP

Abstract

Caveolin-1 is a principal component of caveolae membranes in vivo. Caveolin-1 mRNA and protein expression are lost or reduced during cell transformation by activated oncogenes. Interestingly, the human caveolin-1 gene is localized to a suspected tumor suppressor locus (7q31.1). However, it remains unknown whether downregulation of caveolin-1 is sufficient to mediate cell transformation or tumorigenicity. Here, we employ an antisense approach to derive stable NIH 3T3 cell lines that express dramatically reduced levels of caveolin-1 but contain normal amounts of caveolin-2. NIH 3T3 cells harboring antisense caveolin-1 exhibit anchorage-independent growth, form tumors in immunodeficient mice and show hyperactivation of the p42/44 MAP kinase cascade. Importantly, transformation induced by caveolin-1 downregulation is reversed when caveolin-1 protein levels are restored to normal by loss of the caveolin-1 antisense vector. In addition, we show that in normal NIH 3T3 cells, caveolin-1 expression levels are tightly regulated by specific growth factor stimuli and cell density. Our results suggest that upregulation of caveolin-1 may be important in mediating contact inhibition and negatively regulating the activation state of the p42/44 MAP kinase cascade.

MeSH Terms
3T3 Cells Animals Base Sequence Calcium-Calmodulin-Dependent Protein Kinases/metabolism Caveolin 1 Caveolin 2 Caveolins Cell Transformation, Neoplastic Chromosomes, Human, Pair 7 Culture Media, Serum-Free DNA, Antisense Down-Regulation Enzyme Activation Humans Membrane Proteins/genetics,metabolism Mice Mice, Inbred BALB C Mice, Nude Microscopy, Electron, Scanning Neoplasms, Experimental/genetics Oligonucleotide Probes RNA, Messenger/genetics
Chemicals
CAV1 protein, human Cav1 protein, mouse Caveolin 1 Caveolin 2 Caveolins Culture Media, Serum-Free DNA, Antisense Membrane Proteins Oligonucleotide Probes RNA, Messenger Calcium-Calmodulin-Dependent Protein Kinases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Galbiati F
The Albert Einstein Cancer Center, Microbiology and Immunology, and Epidemiology and Social Medicine, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461, USA.
Volonte D
Engelman J A
Watanabe G
Burk R
Pestell R G
Lisanti M P
References (62)
62 references, click to expand
  1. VIP21, a 21-kD membrane protein is an integral component of trans-Golgi-network-derived transport vesicles.
    J Cell Biol. 1992 Sep;118(5):1003-14 PMID: 1512286
  2. Allelic loss on chromosome 7q in ovarian adenocarcinomas: two critical regions and a rearrangement of the PLANH1 locus.
    Oncogene. 1996 Oct 17;13(8):1815-8 PMID: 8895529
  3. Fish mapping of YAC clones at human chromosomal band 7q31.2: identification of YACS spanning FRA7G within the common region of LOH in breast and prostate cancer.
    Genes Chromosomes Cancer. 1998 Feb;21(2):152-9 PMID: 9491327
  4. Caveolins, a family of scaffolding proteins for organizing "preassembled signaling complexes" at the plasma membrane.
    J Biol Chem. 1998 Mar 6;273(10):5419-22 PMID: 9488658
  5. Tumor cell growth inhibition by caveolin re-expression in human breast cancer cells.
    Oncogene. 1998 Mar;16(11):1391-7 PMID: 9525738
  6. Characterization of caveolin-rich membrane domains isolated from an endothelial-rich source: implications for human disease.
    J Cell Biol. 1994 Jul;126(1):111-26 PMID: 7517942
  7. Reduction of caveolin and caveolae in oncogenically transformed cells.
    Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1381-5 PMID: 7877987
  8. Caveolin interaction with protein kinase C. Isoenzyme-dependent regulation of kinase activity by the caveolin scaffolding domain peptide.
    J Biol Chem. 1997 Dec 26;272(52):33416-21 PMID: 9407137
  9. M-caveolin, a muscle-specific caveolin-related protein.
    FEBS Lett. 1995 Nov 27;376(1-2):108-12 PMID: 8521953
  10. Cell-type and tissue-specific expression of caveolin-2. Caveolins 1 and 2 co-localize and form a stable hetero-oligomeric complex in vivo.
    J Biol Chem. 1997 Nov 14;272(46):29337-46 PMID: 9361015
  11. Caveolae: static inpocketings of the plasma membrane, dynamic vesicles or plain artifact?
    J Cell Sci. 1988 Jul;90 ( Pt 3):341-8 PMID: 3075612
  12. Prognostic significance of allelic imbalance of chromosome arms 7q, 8p, 16q, and 18q in stage T3N0M0 prostate cancer.
    Genes Chromosomes Cancer. 1998 Feb;21(2):131-43 PMID: 9491325
  13. Interaction of a receptor tyrosine kinase, EGF-R, with caveolins. Caveolin binding negatively regulates tyrosine and serine/threonine kinase activities.
    J Biol Chem. 1997 Nov 28;272(48):30429-38 PMID: 9374534
  14. Localization of epidermal growth factor-stimulated Ras/Raf-1 interaction to caveolae membrane.
    J Biol Chem. 1996 May 17;271(20):11930-5 PMID: 8662667
  15. Organized endothelial cell surface signal transduction in caveolae distinct from glycosylphosphatidylinositol-anchored protein microdomains.
    J Biol Chem. 1997 Mar 14;272(11):7211-22 PMID: 9054417
  16. Frequent loss of heterozygosity in the region of the D7S523 locus in advanced ovarian cancer.
    Genes Chromosomes Cancer. 1997 May;19(1):1-5 PMID: 9135988
  17. A detergent-free method for purifying caveolae membrane from tissue culture cells.
    Proc Natl Acad Sci U S A. 1995 Oct 24;92(22):10104-8 PMID: 7479734
  18. Caveolin isoforms differ in their N-terminal protein sequence and subcellular distribution. Identification and epitope mapping of an isoform-specific monoclonal antibody probe.
    J Biol Chem. 1995 Jul 7;270(27):16395-401 PMID: 7608210
  19. Mutational analysis of caveolin-induced vesicle formation. Expression of caveolin-1 recruits caveolin-2 to caveolae membranes.
    FEBS Lett. 1998 Aug 28;434(1-2):127-34 PMID: 9738464
  20. Src tyrosine kinases, Galpha subunits, and H-Ras share a common membrane-anchored scaffolding protein, caveolin. Caveolin binding negatively regulates the auto-activation of Src tyrosine kinases.
    J Biol Chem. 1996 Nov 15;271(46):29182-90 PMID: 8910575
  21. Sequence and expression of caveolin, a protein component of caveolae plasma membrane domains phosphorylated on tyrosine in Rous sarcoma virus-transformed fibroblasts.
    Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10517-21 PMID: 1279683
  22. Platelet-derived growth factor activates mitogen-activated protein kinase in isolated caveolae.
    Proc Natl Acad Sci U S A. 1997 Dec 9;94(25):13666-70 PMID: 9391083
  23. Expression of caveolin-3 in skeletal, cardiac, and smooth muscle cells. Caveolin-3 is a component of the sarcolemma and co-fractionates with dystrophin and dystrophin-associated glycoproteins.
    J Biol Chem. 1996 Jun 21;271(25):15160-5 PMID: 8663016
  24. Targeting of nitric oxide synthase to endothelial cell caveolae via palmitoylation: implications for nitric oxide signaling.
    Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6448-53 PMID: 8692835
  25. Direct interaction of endothelial nitric-oxide synthase and caveolin-1 inhibits synthase activity.
    J Biol Chem. 1997 Jul 25;272(30):18522-5 PMID: 9228013
  26. Morphological changes of the 3T3-L1 fibroblast plasma membrane upon differentiation to the adipocyte form.
    J Cell Sci. 1983 May;61:219-30 PMID: 6885939
  27. Identification, sequence, and expression of caveolin-2 defines a caveolin gene family.
    Proc Natl Acad Sci U S A. 1996 Jan 9;93(1):131-5 PMID: 8552590
  28. Oligomeric structure of caveolin: implications for caveolae membrane organization.
    Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9407-11 PMID: 7568142
  29. Cholesterol depletion of caveolae causes hyperactivation of extracellular signal-related kinase (ERK).
    J Biol Chem. 1998 Aug 14;273(33):21099-104 PMID: 9694863
  30. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
    Nucleic Acids Res. 1983 Mar 11;11(5):1475-89 PMID: 6828386
  31. The sequence of human caveolin reveals identity with VIP21, a component of transport vesicles.
    FEBS Lett. 1992 Dec 7;314(1):45-8 PMID: 1360410
  32. Molecular anatomy of chromosome 7q deletions in myeloid neoplasms: evidence for multiple critical loci.
    Proc Natl Acad Sci U S A. 1998 Mar 31;95(7):3781-5 PMID: 9520444
  33. Localization of platelet-derived growth factor-stimulated phosphorylation cascade to caveolae.
    J Biol Chem. 1996 Apr 26;271(17):10299-303 PMID: 8626598
  34. RNA genetics of breast cancer: maspin as paradigm.
    Cold Spring Harb Symp Quant Biol. 1994;59:537-46 PMID: 7587110
  35. Co-purification and direct interaction of Ras with caveolin, an integral membrane protein of caveolae microdomains. Detergent-free purification of caveolae microdomains.
    J Biol Chem. 1996 Apr 19;271(16):9690-7 PMID: 8621645
  36. Induction of caveolin during adipogenesis and association of GLUT4 with caveolin-rich vesicles.
    J Cell Biol. 1994 Dec;127(5):1233-43 PMID: 7962086
  37. Novel susceptibility locus for mouse hepatomas: evidence for a conserved tumor suppressor gene.
    Genome Res. 1996 Nov;6(11):1070-6 PMID: 8938430
  38. VIP21/caveolin is a cholesterol-binding protein.
    Proc Natl Acad Sci U S A. 1995 Oct 24;92(22):10339-43 PMID: 7479780
  39. Evidence for a regulated interaction between heterotrimeric G proteins and caveolin.
    J Biol Chem. 1995 Jun 30;270(26):15693-701 PMID: 7797570
  40. Molecular cloning of caveolin-3, a novel member of the caveolin gene family expressed predominantly in muscle.
    J Biol Chem. 1996 Jan 26;271(4):2255-61 PMID: 8567687
  41. Identification of peptide and protein ligands for the caveolin-scaffolding domain. Implications for the interaction of caveolin with caveolae-associated proteins.
    J Biol Chem. 1997 Mar 7;272(10):6525-33 PMID: 9045678
  42. Two discrete regions of deletion at 7q in uterine leiomyomas.
    Genes Chromosomes Cancer. 1997 Jul;19(3):156-60 PMID: 9218996
  43. Localization of a tumour-suppressor gene associated with human oral cancer on 7q31.1.
    Int J Cancer. 1998 Mar 2;75(5):671-4 PMID: 9495232
  44. A photo-reactive derivative of ganglioside GM1 specifically cross-links VIP21-caveolin on the cell surface.
    FEBS Lett. 1995 Nov 13;375(1-2):11-4 PMID: 7498456
  45. Loss of heterozygosity on murine chromosome 6 in two-stage carcinogenesis: evidence for a conserved tumor suppressor gene.
    Oncogene. 1997 Jan 9;14(1):109-14 PMID: 9010237
  46. Commonly deleted region on the long arm of chromosome 7 in differentiated adenocarcinoma of the stomach.
    Br J Cancer. 1997;76(12):1567-71 PMID: 9413943
  47. Chromosomal localization, genomic organization, and developmental expression of the murine caveolin gene family (Cav-1, -2, and -3). Cav-1 and Cav-2 genes map to a known tumor suppressor locus (6-A2/7q31).
    FEBS Lett. 1998 Jun 16;429(3):330-6 PMID: 9662443
  48. Membrane-associated vesicules in fibroblasts.
    J Ultrastruct Res. 1977 Nov;61(2):215-7 PMID: 915982
  49. Caveolin-mediated regulation of signaling along the p42/44 MAP kinase cascade in vivo. A role for the caveolin-scaffolding domain.
    FEBS Lett. 1998 May 29;428(3):205-11 PMID: 9654135
  50. The primary sequence of murine caveolin reveals a conserved consensus site for phosphorylation by protein kinase C.
    Gene. 1994 Sep 30;147(2):299-300 PMID: 7926819
  51. Caveolar systems and sarcoplasmic reticulum in coronary smooth muscle cells of the mouse.
    J Ultrastruct Res. 1979 Jun;67(3):325-39 PMID: 458928
  52. Tyrosine phosphorylation of a 22-kDa protein is correlated with transformation by Rous sarcoma virus.
    J Biol Chem. 1989 Dec 5;264(34):20163-6 PMID: 2479645
  53. Reciprocal regulation of neu tyrosine kinase activity and caveolin-1 protein expression in vitro and in vivo. Implications for human breast cancer.
    J Biol Chem. 1998 Aug 7;273(32):20448-55 PMID: 9685399
  54. Loss of heterozygosity on the long arm of human chromosome 7 in sporadic renal cell carcinomas.
    Oncogene. 1997 Nov 27;15(22):2727-33 PMID: 9400999
  55. De novo formation of caveolae in lymphocytes by expression of VIP21-caveolin.
    Proc Natl Acad Sci U S A. 1995 Sep 12;92(19):8655-9 PMID: 7567992
  56. Dissecting the interaction between nitric oxide synthase (NOS) and caveolin. Functional significance of the nos caveolin binding domain in vivo.
    J Biol Chem. 1997 Oct 10;272(41):25437-40 PMID: 9325253
  57. Caveolin, a protein component of caveolae membrane coats.
    Cell. 1992 Feb 21;68(4):673-82 PMID: 1739974
  58. Caveolin versus calmodulin. Counterbalancing allosteric modulators of endothelial nitric oxide synthase.
    J Biol Chem. 1997 Oct 10;272(41):25907-12 PMID: 9325323
  59. Caveolae, caveolin and caveolin-rich membrane domains: a signalling hypothesis.
    Trends Cell Biol. 1994 Jul;4(7):231-5 PMID: 14731661
  60. Acylation targets emdothelial nitric-oxide synthase to plasmalemmal caveolae.
    J Biol Chem. 1996 Mar 15;271(11):6518-22 PMID: 8626455
  61. Expression and characterization of recombinant caveolin. Purification by polyhistidine tagging and cholesterol-dependent incorporation into defined lipid membranes.
    J Biol Chem. 1996 Jan 5;271(1):568-73 PMID: 8550621
  62. Recombinant expression of caveolin-1 in oncogenically transformed cells abrogates anchorage-independent growth.
    J Biol Chem. 1997 Jun 27;272(26):16374-81 PMID: 9195944
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/17.22.6633
Published
1998-11-16
Pages
6633-48
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1171009
Subset
IM
Grants
NCI NIH HHS · R01-CA75503 · United States
NCI NIH HHS · R29-CA70897 · United States
NIGMS NIH HHS · T32-GM07288 · United States
External Links
PubMed source DOI: 10.1093/emboj/17.22.6633 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: product@genelibs.com

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