p53 is phosphorylated by CDK7-cyclin H in a p36(MAT1)-dependent manner

Linda J. Ko, Sheau Yann Shieh, Xinbin Chen, Lata Jayaraman, Katsuyuki Tamai, Yoichi Taya, Carol Prives, Zhen Qiang Pan

Research output: Contribution to journalArticle

140 Citations (Scopus)

Abstract

The tumor suppressor protein p53 acts as a transcriptional activator that can mediate cellular responses to DNA damage by inducing apoptosis and cell cycle arrest, p53 is a nuclear phosphoprotein, and phosphorylation has been proposed to be a means by which the activity of p53 is regulated. The cyclin-dependent kinase (CDK)-activating kinase (CAK) was originally identified as a cellular kinase required for the activation of a CDK-cyclin complex, and CAK is comprised of three subunits: CDK7, cyclin H, and p36(MAT1). CAK is part of the transcription factor IIH multiprotein complex, which is required for RNA polymerase II transcription and nucleotide excision repair. Because of the similarities between p53 and CAK in their involvement in the cell cycle, transcription, and repair, we investigated whether p53 could act as a substrate for phosphorylation by CAK. While CDK7-cyclin H is sufficient for phosphorylation of CDK2, we show that p36(MAT1) is required for efficient phosphorylation of p53 by CDK7-cyclin H, suggesting that p36(MAT1) can act as a substrate specificity-determining factor for CDK7- cyclin H. We have mapped a major site of phosphorylation by CAK to Ser-33 of p53 and have demonstrated as well that p53 is phosphorylated at this site in vivo. Both wild-type and tumor-derived mutant p53 proteins are efficiently phosphorylated by CAK. Furthermore, we show that p36 and p53 can interact both in vitro and in vivo. These studies reveal a potential mechanism for coupling the regulation of p53 with DNA repair and the basal transcriptional machinery.

Original languageEnglish (US)
Pages (from-to)7220-7229
Number of pages10
JournalMolecular and Cellular Biology
Volume17
Issue number12
StatePublished - Dec 1997
Externally publishedYes

Fingerprint

Cyclin H
Phosphotransferases
Phosphorylation
DNA Repair
Transcription Factor TFIIH
Tumor Suppressor Protein p53
Multiprotein Complexes
Cyclins
Cyclin-Dependent Kinases
RNA Polymerase II
Phosphoproteins
Mutant Proteins
Substrate Specificity
Cell Cycle Checkpoints
DNA Damage
Cell Cycle
Apoptosis

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Ko, L. J., Shieh, S. Y., Chen, X., Jayaraman, L., Tamai, K., Taya, Y., ... Pan, Z. Q. (1997). p53 is phosphorylated by CDK7-cyclin H in a p36(MAT1)-dependent manner. Molecular and Cellular Biology, 17(12), 7220-7229.

p53 is phosphorylated by CDK7-cyclin H in a p36(MAT1)-dependent manner. / Ko, Linda J.; Shieh, Sheau Yann; Chen, Xinbin; Jayaraman, Lata; Tamai, Katsuyuki; Taya, Yoichi; Prives, Carol; Pan, Zhen Qiang.

In: Molecular and Cellular Biology, Vol. 17, No. 12, 12.1997, p. 7220-7229.

Research output: Contribution to journalArticle

Ko, LJ, Shieh, SY, Chen, X, Jayaraman, L, Tamai, K, Taya, Y, Prives, C & Pan, ZQ 1997, 'p53 is phosphorylated by CDK7-cyclin H in a p36(MAT1)-dependent manner', Molecular and Cellular Biology, vol. 17, no. 12, pp. 7220-7229.
Ko LJ, Shieh SY, Chen X, Jayaraman L, Tamai K, Taya Y et al. p53 is phosphorylated by CDK7-cyclin H in a p36(MAT1)-dependent manner. Molecular and Cellular Biology. 1997 Dec;17(12):7220-7229.
Ko, Linda J. ; Shieh, Sheau Yann ; Chen, Xinbin ; Jayaraman, Lata ; Tamai, Katsuyuki ; Taya, Yoichi ; Prives, Carol ; Pan, Zhen Qiang. / p53 is phosphorylated by CDK7-cyclin H in a p36(MAT1)-dependent manner. In: Molecular and Cellular Biology. 1997 ; Vol. 17, No. 12. pp. 7220-7229.
@article{fba4d5f907204a5092b8b48e75a3a029,
title = "p53 is phosphorylated by CDK7-cyclin H in a p36(MAT1)-dependent manner",
abstract = "The tumor suppressor protein p53 acts as a transcriptional activator that can mediate cellular responses to DNA damage by inducing apoptosis and cell cycle arrest, p53 is a nuclear phosphoprotein, and phosphorylation has been proposed to be a means by which the activity of p53 is regulated. The cyclin-dependent kinase (CDK)-activating kinase (CAK) was originally identified as a cellular kinase required for the activation of a CDK-cyclin complex, and CAK is comprised of three subunits: CDK7, cyclin H, and p36(MAT1). CAK is part of the transcription factor IIH multiprotein complex, which is required for RNA polymerase II transcription and nucleotide excision repair. Because of the similarities between p53 and CAK in their involvement in the cell cycle, transcription, and repair, we investigated whether p53 could act as a substrate for phosphorylation by CAK. While CDK7-cyclin H is sufficient for phosphorylation of CDK2, we show that p36(MAT1) is required for efficient phosphorylation of p53 by CDK7-cyclin H, suggesting that p36(MAT1) can act as a substrate specificity-determining factor for CDK7- cyclin H. We have mapped a major site of phosphorylation by CAK to Ser-33 of p53 and have demonstrated as well that p53 is phosphorylated at this site in vivo. Both wild-type and tumor-derived mutant p53 proteins are efficiently phosphorylated by CAK. Furthermore, we show that p36 and p53 can interact both in vitro and in vivo. These studies reveal a potential mechanism for coupling the regulation of p53 with DNA repair and the basal transcriptional machinery.",
author = "Ko, {Linda J.} and Shieh, {Sheau Yann} and Xinbin Chen and Lata Jayaraman and Katsuyuki Tamai and Yoichi Taya and Carol Prives and Pan, {Zhen Qiang}",
year = "1997",
month = "12",
language = "English (US)",
volume = "17",
pages = "7220--7229",
journal = "Molecular and Cellular Biology",
issn = "0270-7306",
publisher = "American Society for Microbiology",
number = "12",

}

TY - JOUR

T1 - p53 is phosphorylated by CDK7-cyclin H in a p36(MAT1)-dependent manner

AU - Ko, Linda J.

AU - Shieh, Sheau Yann

AU - Chen, Xinbin

AU - Jayaraman, Lata

AU - Tamai, Katsuyuki

AU - Taya, Yoichi

AU - Prives, Carol

AU - Pan, Zhen Qiang

PY - 1997/12

Y1 - 1997/12

N2 - The tumor suppressor protein p53 acts as a transcriptional activator that can mediate cellular responses to DNA damage by inducing apoptosis and cell cycle arrest, p53 is a nuclear phosphoprotein, and phosphorylation has been proposed to be a means by which the activity of p53 is regulated. The cyclin-dependent kinase (CDK)-activating kinase (CAK) was originally identified as a cellular kinase required for the activation of a CDK-cyclin complex, and CAK is comprised of three subunits: CDK7, cyclin H, and p36(MAT1). CAK is part of the transcription factor IIH multiprotein complex, which is required for RNA polymerase II transcription and nucleotide excision repair. Because of the similarities between p53 and CAK in their involvement in the cell cycle, transcription, and repair, we investigated whether p53 could act as a substrate for phosphorylation by CAK. While CDK7-cyclin H is sufficient for phosphorylation of CDK2, we show that p36(MAT1) is required for efficient phosphorylation of p53 by CDK7-cyclin H, suggesting that p36(MAT1) can act as a substrate specificity-determining factor for CDK7- cyclin H. We have mapped a major site of phosphorylation by CAK to Ser-33 of p53 and have demonstrated as well that p53 is phosphorylated at this site in vivo. Both wild-type and tumor-derived mutant p53 proteins are efficiently phosphorylated by CAK. Furthermore, we show that p36 and p53 can interact both in vitro and in vivo. These studies reveal a potential mechanism for coupling the regulation of p53 with DNA repair and the basal transcriptional machinery.

AB - The tumor suppressor protein p53 acts as a transcriptional activator that can mediate cellular responses to DNA damage by inducing apoptosis and cell cycle arrest, p53 is a nuclear phosphoprotein, and phosphorylation has been proposed to be a means by which the activity of p53 is regulated. The cyclin-dependent kinase (CDK)-activating kinase (CAK) was originally identified as a cellular kinase required for the activation of a CDK-cyclin complex, and CAK is comprised of three subunits: CDK7, cyclin H, and p36(MAT1). CAK is part of the transcription factor IIH multiprotein complex, which is required for RNA polymerase II transcription and nucleotide excision repair. Because of the similarities between p53 and CAK in their involvement in the cell cycle, transcription, and repair, we investigated whether p53 could act as a substrate for phosphorylation by CAK. While CDK7-cyclin H is sufficient for phosphorylation of CDK2, we show that p36(MAT1) is required for efficient phosphorylation of p53 by CDK7-cyclin H, suggesting that p36(MAT1) can act as a substrate specificity-determining factor for CDK7- cyclin H. We have mapped a major site of phosphorylation by CAK to Ser-33 of p53 and have demonstrated as well that p53 is phosphorylated at this site in vivo. Both wild-type and tumor-derived mutant p53 proteins are efficiently phosphorylated by CAK. Furthermore, we show that p36 and p53 can interact both in vitro and in vivo. These studies reveal a potential mechanism for coupling the regulation of p53 with DNA repair and the basal transcriptional machinery.

UR - http://www.scopus.com/inward/record.url?scp=0030724673&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030724673&partnerID=8YFLogxK

M3 - Article

C2 - 9372954

AN - SCOPUS:0030724673

VL - 17

SP - 7220

EP - 7229

JO - Molecular and Cellular Biology

JF - Molecular and Cellular Biology

SN - 0270-7306

IS - 12

ER -