Induction of S-phase entry by E2F transcription factors depends on their nuclear localization

Heiko Müller, Maria Cristina Moroni, Elena Vigo, Birgit Otzen Petersen, Jiri Bartek, Kristian Helin

Research output: Contribution to journalArticle

166 Citations (Scopus)

Abstract

The E2F transcription factors are essential for regulating the correct timing of activation of several genes whose products are implicated in cell proliferation and DNA replication. The E2Fs are targets for negative regulation by the retinoblastoma protein family, which includes pRB, p107, and p130, and they are in a pathway that is frequently found altered in human cancers. There are five members of the E2F family, and they can be divided into two functional subgroups. Whereas, upon overexpression, E2F-1, -2, and - 3 induce S phase in quiescent fibroblasts and override G1 arrests mediated by the p16(INK4A) tumor suppressor protein or neutralizing antibodies to cyclin D1, E2F-4 and -5 do not. Using E2F-1 and E2F-4 as representatives of the two subgroups, we showed here, by constructing a set of chimeric proteins, that the amino terminus of E2F-1 is sufficient to confer S-phase- inducing potential as well as the ability to efficiently transactivate an E2F-responsive promoter to E2F-4. We found that the E2F-1 amino terminus directs chimeric proteins to the nucleus. Surprisingly, a short nuclear localization signal derived from simian virus 40 large T antigen could perfectly substitute for the presence of the E2F-1 amino terminus in these assays. Thus, nuclear y localized E2F-4, when overexpressed, displayed biological activities similar to those of E2F-1. Furthermore, we showed that nuclear localization of endogenous E2F-4 is cell cycle regulated, with E2F-4 being nuclear in the G0 and early G1 phases and mainly cytoplasmic after the pRB family members have become phosphorylated. We propose a novel mechanism for the regulation of E2F-dependent transcription in which E2F-1 regulates transcription only from G0 until mid- to late G1 phase whereas E2F-1 is active in late G1 and S phases, until it is inactivated by cyclin A-dependent kinase in late S phase.

Original languageEnglish
Pages (from-to)5508-5520
Number of pages13
JournalMolecular and Cellular Biology
Volume17
Issue number9
Publication statusPublished - Sep 1997

Fingerprint

E2F Transcription Factors
S Phase
G1 Phase
Cyclin-Dependent Kinase Inhibitor p16
Tumor Suppressor Proteins
Cyclin A
Nuclear Localization Signals
Retinoblastoma Protein
Aptitude
Simian virus 40
Cyclin-Dependent Kinases
Viral Tumor Antigens
Cyclin D1
Neutralizing Antibodies
DNA Replication
Transcriptional Activation
Cell Cycle
Proteins
Fibroblasts
Cell Proliferation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Müller, H., Moroni, M. C., Vigo, E., Petersen, B. O., Bartek, J., & Helin, K. (1997). Induction of S-phase entry by E2F transcription factors depends on their nuclear localization. Molecular and Cellular Biology, 17(9), 5508-5520.

Induction of S-phase entry by E2F transcription factors depends on their nuclear localization. / Müller, Heiko; Moroni, Maria Cristina; Vigo, Elena; Petersen, Birgit Otzen; Bartek, Jiri; Helin, Kristian.

In: Molecular and Cellular Biology, Vol. 17, No. 9, 09.1997, p. 5508-5520.

Research output: Contribution to journalArticle

Müller, H, Moroni, MC, Vigo, E, Petersen, BO, Bartek, J & Helin, K 1997, 'Induction of S-phase entry by E2F transcription factors depends on their nuclear localization', Molecular and Cellular Biology, vol. 17, no. 9, pp. 5508-5520.
Müller, Heiko ; Moroni, Maria Cristina ; Vigo, Elena ; Petersen, Birgit Otzen ; Bartek, Jiri ; Helin, Kristian. / Induction of S-phase entry by E2F transcription factors depends on their nuclear localization. In: Molecular and Cellular Biology. 1997 ; Vol. 17, No. 9. pp. 5508-5520.
@article{aeaba08b4a2c4c538f79579c832cc62e,
title = "Induction of S-phase entry by E2F transcription factors depends on their nuclear localization",
abstract = "The E2F transcription factors are essential for regulating the correct timing of activation of several genes whose products are implicated in cell proliferation and DNA replication. The E2Fs are targets for negative regulation by the retinoblastoma protein family, which includes pRB, p107, and p130, and they are in a pathway that is frequently found altered in human cancers. There are five members of the E2F family, and they can be divided into two functional subgroups. Whereas, upon overexpression, E2F-1, -2, and - 3 induce S phase in quiescent fibroblasts and override G1 arrests mediated by the p16(INK4A) tumor suppressor protein or neutralizing antibodies to cyclin D1, E2F-4 and -5 do not. Using E2F-1 and E2F-4 as representatives of the two subgroups, we showed here, by constructing a set of chimeric proteins, that the amino terminus of E2F-1 is sufficient to confer S-phase- inducing potential as well as the ability to efficiently transactivate an E2F-responsive promoter to E2F-4. We found that the E2F-1 amino terminus directs chimeric proteins to the nucleus. Surprisingly, a short nuclear localization signal derived from simian virus 40 large T antigen could perfectly substitute for the presence of the E2F-1 amino terminus in these assays. Thus, nuclear y localized E2F-4, when overexpressed, displayed biological activities similar to those of E2F-1. Furthermore, we showed that nuclear localization of endogenous E2F-4 is cell cycle regulated, with E2F-4 being nuclear in the G0 and early G1 phases and mainly cytoplasmic after the pRB family members have become phosphorylated. We propose a novel mechanism for the regulation of E2F-dependent transcription in which E2F-1 regulates transcription only from G0 until mid- to late G1 phase whereas E2F-1 is active in late G1 and S phases, until it is inactivated by cyclin A-dependent kinase in late S phase.",
author = "Heiko M{\"u}ller and Moroni, {Maria Cristina} and Elena Vigo and Petersen, {Birgit Otzen} and Jiri Bartek and Kristian Helin",
year = "1997",
month = "9",
language = "English",
volume = "17",
pages = "5508--5520",
journal = "Molecular and Cellular Biology",
issn = "0270-7306",
publisher = "American Society for Microbiology",
number = "9",

}

TY - JOUR

T1 - Induction of S-phase entry by E2F transcription factors depends on their nuclear localization

AU - Müller, Heiko

AU - Moroni, Maria Cristina

AU - Vigo, Elena

AU - Petersen, Birgit Otzen

AU - Bartek, Jiri

AU - Helin, Kristian

PY - 1997/9

Y1 - 1997/9

N2 - The E2F transcription factors are essential for regulating the correct timing of activation of several genes whose products are implicated in cell proliferation and DNA replication. The E2Fs are targets for negative regulation by the retinoblastoma protein family, which includes pRB, p107, and p130, and they are in a pathway that is frequently found altered in human cancers. There are five members of the E2F family, and they can be divided into two functional subgroups. Whereas, upon overexpression, E2F-1, -2, and - 3 induce S phase in quiescent fibroblasts and override G1 arrests mediated by the p16(INK4A) tumor suppressor protein or neutralizing antibodies to cyclin D1, E2F-4 and -5 do not. Using E2F-1 and E2F-4 as representatives of the two subgroups, we showed here, by constructing a set of chimeric proteins, that the amino terminus of E2F-1 is sufficient to confer S-phase- inducing potential as well as the ability to efficiently transactivate an E2F-responsive promoter to E2F-4. We found that the E2F-1 amino terminus directs chimeric proteins to the nucleus. Surprisingly, a short nuclear localization signal derived from simian virus 40 large T antigen could perfectly substitute for the presence of the E2F-1 amino terminus in these assays. Thus, nuclear y localized E2F-4, when overexpressed, displayed biological activities similar to those of E2F-1. Furthermore, we showed that nuclear localization of endogenous E2F-4 is cell cycle regulated, with E2F-4 being nuclear in the G0 and early G1 phases and mainly cytoplasmic after the pRB family members have become phosphorylated. We propose a novel mechanism for the regulation of E2F-dependent transcription in which E2F-1 regulates transcription only from G0 until mid- to late G1 phase whereas E2F-1 is active in late G1 and S phases, until it is inactivated by cyclin A-dependent kinase in late S phase.

AB - The E2F transcription factors are essential for regulating the correct timing of activation of several genes whose products are implicated in cell proliferation and DNA replication. The E2Fs are targets for negative regulation by the retinoblastoma protein family, which includes pRB, p107, and p130, and they are in a pathway that is frequently found altered in human cancers. There are five members of the E2F family, and they can be divided into two functional subgroups. Whereas, upon overexpression, E2F-1, -2, and - 3 induce S phase in quiescent fibroblasts and override G1 arrests mediated by the p16(INK4A) tumor suppressor protein or neutralizing antibodies to cyclin D1, E2F-4 and -5 do not. Using E2F-1 and E2F-4 as representatives of the two subgroups, we showed here, by constructing a set of chimeric proteins, that the amino terminus of E2F-1 is sufficient to confer S-phase- inducing potential as well as the ability to efficiently transactivate an E2F-responsive promoter to E2F-4. We found that the E2F-1 amino terminus directs chimeric proteins to the nucleus. Surprisingly, a short nuclear localization signal derived from simian virus 40 large T antigen could perfectly substitute for the presence of the E2F-1 amino terminus in these assays. Thus, nuclear y localized E2F-4, when overexpressed, displayed biological activities similar to those of E2F-1. Furthermore, we showed that nuclear localization of endogenous E2F-4 is cell cycle regulated, with E2F-4 being nuclear in the G0 and early G1 phases and mainly cytoplasmic after the pRB family members have become phosphorylated. We propose a novel mechanism for the regulation of E2F-dependent transcription in which E2F-1 regulates transcription only from G0 until mid- to late G1 phase whereas E2F-1 is active in late G1 and S phases, until it is inactivated by cyclin A-dependent kinase in late S phase.

UR - http://www.scopus.com/inward/record.url?scp=0039227088&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0039227088&partnerID=8YFLogxK

M3 - Article

C2 - 9271426

AN - SCOPUS:0039227088

VL - 17

SP - 5508

EP - 5520

JO - Molecular and Cellular Biology

JF - Molecular and Cellular Biology

SN - 0270-7306

IS - 9

ER -