TGF-β mediated G1 arrest in a human melanoma cell line lacking p15(INK4B): evidence for cooperation between p21(Cip1/WAF1) and p27(Kip1)

V. A. Florenes, N. Bhattacharya, M. R. Bani, Y. Ben-David, R. S. Kerbel, J. M. Slingerland

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

We have studied TGF-β mediated G1 arrest in WM35, an early stage human melanoma cell line. These cells have lost p15(INK4B) expression through loss of one chromosome 9 and rearrangement of the other. In asynchronously growing WM35, TGF-β caused reductions in cyclin D1, cyclin A and cdk4 proteins and their associated kinase activities and an increase in both p21(Cip1/WAF1) and p27(Kip1). These findings were confirmed in cells released from quiescence in the presence of TGF-β, in which TGF-β inhibited or delayed the reduction in the cdk inhibitors that normally occurs in late G1. In contrast to observations in other cell types, there was an increased association of both p21(Cip1/WAF1) and p27(Kip1) with cyclin D1/cdk4 and with cyclin E/cdk2 during TGF-β mediated arrest of asynchronously growing cells. Upregulation of p21(Cip1/WAF1) preceded that of p27(Kip1). Furthermore, p21(Cip1/WAF1) and p27(Kip1) were not present in the same cdk complexes but bound distinct populations of target cdk molecules. Both p21(Cip1/WAF1) and p27(Kip1) immunoprecipitates from asynchronously growing cells contained active kinase complexes. These KIP-associated kinase activities were reduced in TGF-β arrested cells. It has been proposed that in TGF-β arrested epithelial cells, up-regulation of p15(INK4B) and of p15(INK4B) binding to cdk4 serves to destabilize the association of p27(Kip1) With cyclin D1/cdk4, promoting p27(Kip1) binding and inhibition of cyclin E/cdk2. Our findings demonstrate that this is not a universal mechanism of G1 arrest by TGF-β. In TGF-β arrested WM35, which lack p15(INK4B), the increased p21(Cip1/WAF1) may serve a similar function to that of p15(INK4B): initiating kinase inhibition and providing an additional mechanism to supplement the effect of p27(KiP1) on G1 cyclin/cdks.

Original languageEnglish
Pages (from-to)2447-2457
Number of pages11
JournalOncogene
Volume13
Issue number11
Publication statusPublished - 1996

Fingerprint

Melanoma
Cell Line
Cyclin D1
Phosphotransferases
Cyclin E
Up-Regulation
Cyclin G1
Cyclin-Dependent Kinase 4
Cyclin A
Chromosomes, Human, Pair 9
Health Services Needs and Demand
Epithelial Cells

Keywords

  • G arrest
  • Melanoma
  • p15(INK4B)-negative
  • p21(Cip1/WAF1)
  • p27(Kip1)
  • TFG-β

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Florenes, V. A., Bhattacharya, N., Bani, M. R., Ben-David, Y., Kerbel, R. S., & Slingerland, J. M. (1996). TGF-β mediated G1 arrest in a human melanoma cell line lacking p15(INK4B): evidence for cooperation between p21(Cip1/WAF1) and p27(Kip1). Oncogene, 13(11), 2447-2457.

TGF-β mediated G1 arrest in a human melanoma cell line lacking p15(INK4B) : evidence for cooperation between p21(Cip1/WAF1) and p27(Kip1). / Florenes, V. A.; Bhattacharya, N.; Bani, M. R.; Ben-David, Y.; Kerbel, R. S.; Slingerland, J. M.

In: Oncogene, Vol. 13, No. 11, 1996, p. 2447-2457.

Research output: Contribution to journalArticle

Florenes, VA, Bhattacharya, N, Bani, MR, Ben-David, Y, Kerbel, RS & Slingerland, JM 1996, 'TGF-β mediated G1 arrest in a human melanoma cell line lacking p15(INK4B): evidence for cooperation between p21(Cip1/WAF1) and p27(Kip1)', Oncogene, vol. 13, no. 11, pp. 2447-2457.
Florenes, V. A. ; Bhattacharya, N. ; Bani, M. R. ; Ben-David, Y. ; Kerbel, R. S. ; Slingerland, J. M. / TGF-β mediated G1 arrest in a human melanoma cell line lacking p15(INK4B) : evidence for cooperation between p21(Cip1/WAF1) and p27(Kip1). In: Oncogene. 1996 ; Vol. 13, No. 11. pp. 2447-2457.
@article{46e40a25a07041e98b0aaabf0b56ed5f,
title = "TGF-β mediated G1 arrest in a human melanoma cell line lacking p15(INK4B): evidence for cooperation between p21(Cip1/WAF1) and p27(Kip1)",
abstract = "We have studied TGF-β mediated G1 arrest in WM35, an early stage human melanoma cell line. These cells have lost p15(INK4B) expression through loss of one chromosome 9 and rearrangement of the other. In asynchronously growing WM35, TGF-β caused reductions in cyclin D1, cyclin A and cdk4 proteins and their associated kinase activities and an increase in both p21(Cip1/WAF1) and p27(Kip1). These findings were confirmed in cells released from quiescence in the presence of TGF-β, in which TGF-β inhibited or delayed the reduction in the cdk inhibitors that normally occurs in late G1. In contrast to observations in other cell types, there was an increased association of both p21(Cip1/WAF1) and p27(Kip1) with cyclin D1/cdk4 and with cyclin E/cdk2 during TGF-β mediated arrest of asynchronously growing cells. Upregulation of p21(Cip1/WAF1) preceded that of p27(Kip1). Furthermore, p21(Cip1/WAF1) and p27(Kip1) were not present in the same cdk complexes but bound distinct populations of target cdk molecules. Both p21(Cip1/WAF1) and p27(Kip1) immunoprecipitates from asynchronously growing cells contained active kinase complexes. These KIP-associated kinase activities were reduced in TGF-β arrested cells. It has been proposed that in TGF-β arrested epithelial cells, up-regulation of p15(INK4B) and of p15(INK4B) binding to cdk4 serves to destabilize the association of p27(Kip1) With cyclin D1/cdk4, promoting p27(Kip1) binding and inhibition of cyclin E/cdk2. Our findings demonstrate that this is not a universal mechanism of G1 arrest by TGF-β. In TGF-β arrested WM35, which lack p15(INK4B), the increased p21(Cip1/WAF1) may serve a similar function to that of p15(INK4B): initiating kinase inhibition and providing an additional mechanism to supplement the effect of p27(KiP1) on G1 cyclin/cdks.",
keywords = "G arrest, Melanoma, p15(INK4B)-negative, p21(Cip1/WAF1), p27(Kip1), TFG-β",
author = "Florenes, {V. A.} and N. Bhattacharya and Bani, {M. R.} and Y. Ben-David and Kerbel, {R. S.} and Slingerland, {J. M.}",
year = "1996",
language = "English",
volume = "13",
pages = "2447--2457",
journal = "Oncogene",
issn = "0950-9232",
publisher = "Nature Publishing Group",
number = "11",

}

TY - JOUR

T1 - TGF-β mediated G1 arrest in a human melanoma cell line lacking p15(INK4B)

T2 - evidence for cooperation between p21(Cip1/WAF1) and p27(Kip1)

AU - Florenes, V. A.

AU - Bhattacharya, N.

AU - Bani, M. R.

AU - Ben-David, Y.

AU - Kerbel, R. S.

AU - Slingerland, J. M.

PY - 1996

Y1 - 1996

N2 - We have studied TGF-β mediated G1 arrest in WM35, an early stage human melanoma cell line. These cells have lost p15(INK4B) expression through loss of one chromosome 9 and rearrangement of the other. In asynchronously growing WM35, TGF-β caused reductions in cyclin D1, cyclin A and cdk4 proteins and their associated kinase activities and an increase in both p21(Cip1/WAF1) and p27(Kip1). These findings were confirmed in cells released from quiescence in the presence of TGF-β, in which TGF-β inhibited or delayed the reduction in the cdk inhibitors that normally occurs in late G1. In contrast to observations in other cell types, there was an increased association of both p21(Cip1/WAF1) and p27(Kip1) with cyclin D1/cdk4 and with cyclin E/cdk2 during TGF-β mediated arrest of asynchronously growing cells. Upregulation of p21(Cip1/WAF1) preceded that of p27(Kip1). Furthermore, p21(Cip1/WAF1) and p27(Kip1) were not present in the same cdk complexes but bound distinct populations of target cdk molecules. Both p21(Cip1/WAF1) and p27(Kip1) immunoprecipitates from asynchronously growing cells contained active kinase complexes. These KIP-associated kinase activities were reduced in TGF-β arrested cells. It has been proposed that in TGF-β arrested epithelial cells, up-regulation of p15(INK4B) and of p15(INK4B) binding to cdk4 serves to destabilize the association of p27(Kip1) With cyclin D1/cdk4, promoting p27(Kip1) binding and inhibition of cyclin E/cdk2. Our findings demonstrate that this is not a universal mechanism of G1 arrest by TGF-β. In TGF-β arrested WM35, which lack p15(INK4B), the increased p21(Cip1/WAF1) may serve a similar function to that of p15(INK4B): initiating kinase inhibition and providing an additional mechanism to supplement the effect of p27(KiP1) on G1 cyclin/cdks.

AB - We have studied TGF-β mediated G1 arrest in WM35, an early stage human melanoma cell line. These cells have lost p15(INK4B) expression through loss of one chromosome 9 and rearrangement of the other. In asynchronously growing WM35, TGF-β caused reductions in cyclin D1, cyclin A and cdk4 proteins and their associated kinase activities and an increase in both p21(Cip1/WAF1) and p27(Kip1). These findings were confirmed in cells released from quiescence in the presence of TGF-β, in which TGF-β inhibited or delayed the reduction in the cdk inhibitors that normally occurs in late G1. In contrast to observations in other cell types, there was an increased association of both p21(Cip1/WAF1) and p27(Kip1) with cyclin D1/cdk4 and with cyclin E/cdk2 during TGF-β mediated arrest of asynchronously growing cells. Upregulation of p21(Cip1/WAF1) preceded that of p27(Kip1). Furthermore, p21(Cip1/WAF1) and p27(Kip1) were not present in the same cdk complexes but bound distinct populations of target cdk molecules. Both p21(Cip1/WAF1) and p27(Kip1) immunoprecipitates from asynchronously growing cells contained active kinase complexes. These KIP-associated kinase activities were reduced in TGF-β arrested cells. It has been proposed that in TGF-β arrested epithelial cells, up-regulation of p15(INK4B) and of p15(INK4B) binding to cdk4 serves to destabilize the association of p27(Kip1) With cyclin D1/cdk4, promoting p27(Kip1) binding and inhibition of cyclin E/cdk2. Our findings demonstrate that this is not a universal mechanism of G1 arrest by TGF-β. In TGF-β arrested WM35, which lack p15(INK4B), the increased p21(Cip1/WAF1) may serve a similar function to that of p15(INK4B): initiating kinase inhibition and providing an additional mechanism to supplement the effect of p27(KiP1) on G1 cyclin/cdks.

KW - G arrest

KW - Melanoma

KW - p15(INK4B)-negative

KW - p21(Cip1/WAF1)

KW - p27(Kip1)

KW - TFG-β

UR - http://www.scopus.com/inward/record.url?scp=0030457645&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030457645&partnerID=8YFLogxK

M3 - Article

C2 - 8957087

AN - SCOPUS:0030457645

VL - 13

SP - 2447

EP - 2457

JO - Oncogene

JF - Oncogene

SN - 0950-9232

IS - 11

ER -