Iron deprivation inhibits cyclin-dependent kinase activity and decreases cyclin D/CDK4 protein levels in asynchronous MDA-MB-453 human breast cancer cells

Kristen S. Kulp, Sherril L. Green, Philip R Vulliet

Research output: Contribution to journalArticle

87 Citations (Scopus)

Abstract

Iron chelation, known to block progression through the cell cycle, was examined for effects on the activity and subunit levels of the cyclin- dependent protein kinases (cdk). Treatment of asynchronous MDA-MB-453 cells with the iron chelators mimosine or desferrioxamine (DFO) for 24 h stopped cell division, but did not produce a single, synchronous block. DNA content analysis demonstrated that although a majority of the cells were blocked in G1 (87.3%), an unexpectedly large fraction of the cells were blocked in S phase (11.5%). Western blot analysis of the treated lysates demonstrated the presence of cyclin B, confirming that part of the cell population was blocked in S phase. After release from mimosine treatment, 84% of the cell population remained in G1 up to 8 h. Treating breast cancer cells with 400 ηM mimosine for 24 h inhibited cyclin E- and cyclin A-associated kinase activity by 85% or more, although immunoblots using anti-cyclin A, cyclin E, cdc2, and cdk2 antibodies showed that these key subunits were still present in the cells at pretreatment levels. Interestingly, Western blot analysis also demonstrated that iron chelation decreased the protein levels of the cyclin D and cdk4 subunits as compared to control and produced a change in retinoblastoma protein phosphorylation. These results indicate that iron deprivation effects the activity and protein levels of the cyclin-dependent kinases, and ultimately, the pathways that control cell division.

Original languageEnglish (US)
Pages (from-to)60-68
Number of pages9
JournalExperimental Cell Research
Volume229
Issue number1
DOIs
StatePublished - Nov 25 1996

Fingerprint

Cyclin D
Cyclin-Dependent Kinases
Iron
Breast Neoplasms
Mimosine
Proteins
Cyclin A
Cyclin E
S Phase
Cell Division
Western Blotting
Cyclin B
Retinoblastoma Protein
Deferoxamine
Chelating Agents
Population
Cell Cycle
Phosphotransferases
Phosphorylation
Antibodies

ASJC Scopus subject areas

  • Cell Biology

Cite this

Iron deprivation inhibits cyclin-dependent kinase activity and decreases cyclin D/CDK4 protein levels in asynchronous MDA-MB-453 human breast cancer cells. / Kulp, Kristen S.; Green, Sherril L.; Vulliet, Philip R.

In: Experimental Cell Research, Vol. 229, No. 1, 25.11.1996, p. 60-68.

Research output: Contribution to journalArticle

@article{9325efae2e4c4bbfbcbb58ce1225c14b,
title = "Iron deprivation inhibits cyclin-dependent kinase activity and decreases cyclin D/CDK4 protein levels in asynchronous MDA-MB-453 human breast cancer cells",
abstract = "Iron chelation, known to block progression through the cell cycle, was examined for effects on the activity and subunit levels of the cyclin- dependent protein kinases (cdk). Treatment of asynchronous MDA-MB-453 cells with the iron chelators mimosine or desferrioxamine (DFO) for 24 h stopped cell division, but did not produce a single, synchronous block. DNA content analysis demonstrated that although a majority of the cells were blocked in G1 (87.3{\%}), an unexpectedly large fraction of the cells were blocked in S phase (11.5{\%}). Western blot analysis of the treated lysates demonstrated the presence of cyclin B, confirming that part of the cell population was blocked in S phase. After release from mimosine treatment, 84{\%} of the cell population remained in G1 up to 8 h. Treating breast cancer cells with 400 ηM mimosine for 24 h inhibited cyclin E- and cyclin A-associated kinase activity by 85{\%} or more, although immunoblots using anti-cyclin A, cyclin E, cdc2, and cdk2 antibodies showed that these key subunits were still present in the cells at pretreatment levels. Interestingly, Western blot analysis also demonstrated that iron chelation decreased the protein levels of the cyclin D and cdk4 subunits as compared to control and produced a change in retinoblastoma protein phosphorylation. These results indicate that iron deprivation effects the activity and protein levels of the cyclin-dependent kinases, and ultimately, the pathways that control cell division.",
author = "Kulp, {Kristen S.} and Green, {Sherril L.} and Vulliet, {Philip R}",
year = "1996",
month = "11",
day = "25",
doi = "10.1006/excr.1996.0343",
language = "English (US)",
volume = "229",
pages = "60--68",
journal = "Experimental Cell Research",
issn = "0014-4827",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Iron deprivation inhibits cyclin-dependent kinase activity and decreases cyclin D/CDK4 protein levels in asynchronous MDA-MB-453 human breast cancer cells

AU - Kulp, Kristen S.

AU - Green, Sherril L.

AU - Vulliet, Philip R

PY - 1996/11/25

Y1 - 1996/11/25

N2 - Iron chelation, known to block progression through the cell cycle, was examined for effects on the activity and subunit levels of the cyclin- dependent protein kinases (cdk). Treatment of asynchronous MDA-MB-453 cells with the iron chelators mimosine or desferrioxamine (DFO) for 24 h stopped cell division, but did not produce a single, synchronous block. DNA content analysis demonstrated that although a majority of the cells were blocked in G1 (87.3%), an unexpectedly large fraction of the cells were blocked in S phase (11.5%). Western blot analysis of the treated lysates demonstrated the presence of cyclin B, confirming that part of the cell population was blocked in S phase. After release from mimosine treatment, 84% of the cell population remained in G1 up to 8 h. Treating breast cancer cells with 400 ηM mimosine for 24 h inhibited cyclin E- and cyclin A-associated kinase activity by 85% or more, although immunoblots using anti-cyclin A, cyclin E, cdc2, and cdk2 antibodies showed that these key subunits were still present in the cells at pretreatment levels. Interestingly, Western blot analysis also demonstrated that iron chelation decreased the protein levels of the cyclin D and cdk4 subunits as compared to control and produced a change in retinoblastoma protein phosphorylation. These results indicate that iron deprivation effects the activity and protein levels of the cyclin-dependent kinases, and ultimately, the pathways that control cell division.

AB - Iron chelation, known to block progression through the cell cycle, was examined for effects on the activity and subunit levels of the cyclin- dependent protein kinases (cdk). Treatment of asynchronous MDA-MB-453 cells with the iron chelators mimosine or desferrioxamine (DFO) for 24 h stopped cell division, but did not produce a single, synchronous block. DNA content analysis demonstrated that although a majority of the cells were blocked in G1 (87.3%), an unexpectedly large fraction of the cells were blocked in S phase (11.5%). Western blot analysis of the treated lysates demonstrated the presence of cyclin B, confirming that part of the cell population was blocked in S phase. After release from mimosine treatment, 84% of the cell population remained in G1 up to 8 h. Treating breast cancer cells with 400 ηM mimosine for 24 h inhibited cyclin E- and cyclin A-associated kinase activity by 85% or more, although immunoblots using anti-cyclin A, cyclin E, cdc2, and cdk2 antibodies showed that these key subunits were still present in the cells at pretreatment levels. Interestingly, Western blot analysis also demonstrated that iron chelation decreased the protein levels of the cyclin D and cdk4 subunits as compared to control and produced a change in retinoblastoma protein phosphorylation. These results indicate that iron deprivation effects the activity and protein levels of the cyclin-dependent kinases, and ultimately, the pathways that control cell division.

UR - http://www.scopus.com/inward/record.url?scp=0030601987&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030601987&partnerID=8YFLogxK

U2 - 10.1006/excr.1996.0343

DO - 10.1006/excr.1996.0343

M3 - Article

C2 - 8940249

AN - SCOPUS:0030601987

VL - 229

SP - 60

EP - 68

JO - Experimental Cell Research

JF - Experimental Cell Research

SN - 0014-4827

IS - 1

ER -