Von Hippel-Lindau protein complex is regulated by cell density

Sankar Mohan, Robert D. Burk

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Mutations in the von Hippel-Lindau (VHL) gene are involved in the VHL family cancer syndrome and sporadic renal cell carcinoma. Previous studies indicated that VHL-induced growth arrest required high cell density and growth on extracellular matrix. In the present study, VHL protein (pVHL) levels were observed to be dramatically increased in cells grown to high cell density compared to cells grown at low cell density. Reverse transcription-polymerase chain reaction and Northern blot analysis indicated that VHL mRNA levels were equivalent in sparse and dense cells. The pVHL was rapidly degraded when cell-cell contact was disturbed by trypsinization or EDTA release. Treatment of cells with a proteasome inhibitor blocked the degradation of pVHL. Using a set of VHL deletions fused to GFP, a cell density-dependent region (CDDR) was identified and localized to the c-terminus of pVHL. In addition, other members of the VBC protein complex also showed a cell density-dependent regulation similar to pVHL. Cell density regulation of VHL did not require elongin binding and density-dependent regulation of other VBC components was not dependent on pVHL. In addition, hypoxia inducible factor-2α protein levels were elevated in sparse cells with low levels of pVHL and reduced or absent in confluent cells containing abundant VHL. These results indicate that pVHL levels and thus function are tightly regulated by cell-cell signaling. In addition, care must be taken when interpreting studies of VHL function and subcellular localization of cells grown at subconfluent conditions.

Original languageEnglish (US)
Pages (from-to)5270-5280
Number of pages11
JournalOncogene
Volume22
Issue number34
DOIs
StatePublished - Aug 14 2003

Fingerprint

Cell Count
Proteins
Proteasome Inhibitors
Growth
Renal Cell Carcinoma
Edetic Acid
Northern Blotting
Reverse Transcription
Extracellular Matrix
Polymerase Chain Reaction
Messenger RNA
Mutation
Genes

Keywords

  • Cell density
  • Cell-cell contact
  • VHL

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Von Hippel-Lindau protein complex is regulated by cell density. / Mohan, Sankar; Burk, Robert D.

In: Oncogene, Vol. 22, No. 34, 14.08.2003, p. 5270-5280.

Research output: Contribution to journalArticle

Mohan, Sankar ; Burk, Robert D. / Von Hippel-Lindau protein complex is regulated by cell density. In: Oncogene. 2003 ; Vol. 22, No. 34. pp. 5270-5280.
@article{c71027c6ebb94ec6be48b0e41ba580e4,
title = "Von Hippel-Lindau protein complex is regulated by cell density",
abstract = "Mutations in the von Hippel-Lindau (VHL) gene are involved in the VHL family cancer syndrome and sporadic renal cell carcinoma. Previous studies indicated that VHL-induced growth arrest required high cell density and growth on extracellular matrix. In the present study, VHL protein (pVHL) levels were observed to be dramatically increased in cells grown to high cell density compared to cells grown at low cell density. Reverse transcription-polymerase chain reaction and Northern blot analysis indicated that VHL mRNA levels were equivalent in sparse and dense cells. The pVHL was rapidly degraded when cell-cell contact was disturbed by trypsinization or EDTA release. Treatment of cells with a proteasome inhibitor blocked the degradation of pVHL. Using a set of VHL deletions fused to GFP, a cell density-dependent region (CDDR) was identified and localized to the c-terminus of pVHL. In addition, other members of the VBC protein complex also showed a cell density-dependent regulation similar to pVHL. Cell density regulation of VHL did not require elongin binding and density-dependent regulation of other VBC components was not dependent on pVHL. In addition, hypoxia inducible factor-2α protein levels were elevated in sparse cells with low levels of pVHL and reduced or absent in confluent cells containing abundant VHL. These results indicate that pVHL levels and thus function are tightly regulated by cell-cell signaling. In addition, care must be taken when interpreting studies of VHL function and subcellular localization of cells grown at subconfluent conditions.",
keywords = "Cell density, Cell-cell contact, VHL",
author = "Sankar Mohan and Burk, {Robert D.}",
year = "2003",
month = "8",
day = "14",
doi = "10.1038/sj.onc.1206592",
language = "English (US)",
volume = "22",
pages = "5270--5280",
journal = "Oncogene",
issn = "0950-9232",
publisher = "Nature Publishing Group",
number = "34",

}

TY - JOUR

T1 - Von Hippel-Lindau protein complex is regulated by cell density

AU - Mohan, Sankar

AU - Burk, Robert D.

PY - 2003/8/14

Y1 - 2003/8/14

N2 - Mutations in the von Hippel-Lindau (VHL) gene are involved in the VHL family cancer syndrome and sporadic renal cell carcinoma. Previous studies indicated that VHL-induced growth arrest required high cell density and growth on extracellular matrix. In the present study, VHL protein (pVHL) levels were observed to be dramatically increased in cells grown to high cell density compared to cells grown at low cell density. Reverse transcription-polymerase chain reaction and Northern blot analysis indicated that VHL mRNA levels were equivalent in sparse and dense cells. The pVHL was rapidly degraded when cell-cell contact was disturbed by trypsinization or EDTA release. Treatment of cells with a proteasome inhibitor blocked the degradation of pVHL. Using a set of VHL deletions fused to GFP, a cell density-dependent region (CDDR) was identified and localized to the c-terminus of pVHL. In addition, other members of the VBC protein complex also showed a cell density-dependent regulation similar to pVHL. Cell density regulation of VHL did not require elongin binding and density-dependent regulation of other VBC components was not dependent on pVHL. In addition, hypoxia inducible factor-2α protein levels were elevated in sparse cells with low levels of pVHL and reduced or absent in confluent cells containing abundant VHL. These results indicate that pVHL levels and thus function are tightly regulated by cell-cell signaling. In addition, care must be taken when interpreting studies of VHL function and subcellular localization of cells grown at subconfluent conditions.

AB - Mutations in the von Hippel-Lindau (VHL) gene are involved in the VHL family cancer syndrome and sporadic renal cell carcinoma. Previous studies indicated that VHL-induced growth arrest required high cell density and growth on extracellular matrix. In the present study, VHL protein (pVHL) levels were observed to be dramatically increased in cells grown to high cell density compared to cells grown at low cell density. Reverse transcription-polymerase chain reaction and Northern blot analysis indicated that VHL mRNA levels were equivalent in sparse and dense cells. The pVHL was rapidly degraded when cell-cell contact was disturbed by trypsinization or EDTA release. Treatment of cells with a proteasome inhibitor blocked the degradation of pVHL. Using a set of VHL deletions fused to GFP, a cell density-dependent region (CDDR) was identified and localized to the c-terminus of pVHL. In addition, other members of the VBC protein complex also showed a cell density-dependent regulation similar to pVHL. Cell density regulation of VHL did not require elongin binding and density-dependent regulation of other VBC components was not dependent on pVHL. In addition, hypoxia inducible factor-2α protein levels were elevated in sparse cells with low levels of pVHL and reduced or absent in confluent cells containing abundant VHL. These results indicate that pVHL levels and thus function are tightly regulated by cell-cell signaling. In addition, care must be taken when interpreting studies of VHL function and subcellular localization of cells grown at subconfluent conditions.

KW - Cell density

KW - Cell-cell contact

KW - VHL

UR - http://www.scopus.com/inward/record.url?scp=0041353455&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0041353455&partnerID=8YFLogxK

U2 - 10.1038/sj.onc.1206592

DO - 10.1038/sj.onc.1206592

M3 - Article

C2 - 12917628

AN - SCOPUS:0041353455

VL - 22

SP - 5270

EP - 5280

JO - Oncogene

JF - Oncogene

SN - 0950-9232

IS - 34

ER -