Expression cloning of a cDNA encoding a retinoblastoma-binding protein with E2F-like properties

William G. Kaelin, Wilhelm Krek, William R. Sellers, James A. DeCaprio, Florence Ajchenbaum, Charles S. Fuchs, Thomas Chittenden, Yue Li, Peggy J. Farnham, Michael A. Blanar, David M. Livingston, Erik K. Flemington

Research output: Contribution to journalArticle

655 Citations (Scopus)

Abstract

An expression vector was modified to permit the rapid synthesis of purified, 32P-labeled, glutathione S-transferase (GST)-retinoblastoma (RB) fusion proteins. The products were used to screen λgt11 expression libraries, from which we cloned a cDNA encoding a polypeptide (RBAP-1) capable of binding directly to a putative functional domain (the pocket) of the retinoblastoma gene product (RB). The RB "pocket" is known to bind, directly or indirectly, to the cellular transcription factor, E2F, implicated in cell growth control. We have found that RBAP-1 copurifies with E2F, interacts specifically with the adenovirus E4 ORF 6 7 protein, binds specifically and directly to a known E2F DNA recognition sequence, and contains a functional transactivation domain. Therefore, RBAP-1 is a species of E2F and can bind specifically to the RB pocket.

Original languageEnglish (US)
Pages (from-to)351-364
Number of pages14
JournalCell
Volume70
Issue number2
DOIs
StatePublished - Jul 24 1992
Externally publishedYes

Fingerprint

Retinoblastoma Binding Proteins
Retinoblastoma
Cloning
Organism Cloning
Complementary DNA
E2F Transcription Factors
Cell growth
Glutathione Transferase
Retinoblastoma Genes
Retinoblastoma Protein
Proteins
Fusion reactions
Genes
Adenoviridae
Peptides
Transcriptional Activation
Open Reading Frames
DNA
Growth

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology

Cite this

Kaelin, W. G., Krek, W., Sellers, W. R., DeCaprio, J. A., Ajchenbaum, F., Fuchs, C. S., ... Flemington, E. K. (1992). Expression cloning of a cDNA encoding a retinoblastoma-binding protein with E2F-like properties. Cell, 70(2), 351-364. https://doi.org/10.1016/0092-8674(92)90108-O

Expression cloning of a cDNA encoding a retinoblastoma-binding protein with E2F-like properties. / Kaelin, William G.; Krek, Wilhelm; Sellers, William R.; DeCaprio, James A.; Ajchenbaum, Florence; Fuchs, Charles S.; Chittenden, Thomas; Li, Yue; Farnham, Peggy J.; Blanar, Michael A.; Livingston, David M.; Flemington, Erik K.

In: Cell, Vol. 70, No. 2, 24.07.1992, p. 351-364.

Research output: Contribution to journalArticle

Kaelin, WG, Krek, W, Sellers, WR, DeCaprio, JA, Ajchenbaum, F, Fuchs, CS, Chittenden, T, Li, Y, Farnham, PJ, Blanar, MA, Livingston, DM & Flemington, EK 1992, 'Expression cloning of a cDNA encoding a retinoblastoma-binding protein with E2F-like properties', Cell, vol. 70, no. 2, pp. 351-364. https://doi.org/10.1016/0092-8674(92)90108-O
Kaelin WG, Krek W, Sellers WR, DeCaprio JA, Ajchenbaum F, Fuchs CS et al. Expression cloning of a cDNA encoding a retinoblastoma-binding protein with E2F-like properties. Cell. 1992 Jul 24;70(2):351-364. https://doi.org/10.1016/0092-8674(92)90108-O
Kaelin, William G. ; Krek, Wilhelm ; Sellers, William R. ; DeCaprio, James A. ; Ajchenbaum, Florence ; Fuchs, Charles S. ; Chittenden, Thomas ; Li, Yue ; Farnham, Peggy J. ; Blanar, Michael A. ; Livingston, David M. ; Flemington, Erik K. / Expression cloning of a cDNA encoding a retinoblastoma-binding protein with E2F-like properties. In: Cell. 1992 ; Vol. 70, No. 2. pp. 351-364.
@article{7222072c36e646e786dc2cb8fbe66eea,
title = "Expression cloning of a cDNA encoding a retinoblastoma-binding protein with E2F-like properties",
abstract = "An expression vector was modified to permit the rapid synthesis of purified, 32P-labeled, glutathione S-transferase (GST)-retinoblastoma (RB) fusion proteins. The products were used to screen λgt11 expression libraries, from which we cloned a cDNA encoding a polypeptide (RBAP-1) capable of binding directly to a putative functional domain (the pocket) of the retinoblastoma gene product (RB). The RB {"}pocket{"} is known to bind, directly or indirectly, to the cellular transcription factor, E2F, implicated in cell growth control. We have found that RBAP-1 copurifies with E2F, interacts specifically with the adenovirus E4 ORF 6 7 protein, binds specifically and directly to a known E2F DNA recognition sequence, and contains a functional transactivation domain. Therefore, RBAP-1 is a species of E2F and can bind specifically to the RB pocket.",
author = "Kaelin, {William G.} and Wilhelm Krek and Sellers, {William R.} and DeCaprio, {James A.} and Florence Ajchenbaum and Fuchs, {Charles S.} and Thomas Chittenden and Yue Li and Farnham, {Peggy J.} and Blanar, {Michael A.} and Livingston, {David M.} and Flemington, {Erik K.}",
year = "1992",
month = "7",
day = "24",
doi = "10.1016/0092-8674(92)90108-O",
language = "English (US)",
volume = "70",
pages = "351--364",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "2",

}

TY - JOUR

T1 - Expression cloning of a cDNA encoding a retinoblastoma-binding protein with E2F-like properties

AU - Kaelin, William G.

AU - Krek, Wilhelm

AU - Sellers, William R.

AU - DeCaprio, James A.

AU - Ajchenbaum, Florence

AU - Fuchs, Charles S.

AU - Chittenden, Thomas

AU - Li, Yue

AU - Farnham, Peggy J.

AU - Blanar, Michael A.

AU - Livingston, David M.

AU - Flemington, Erik K.

PY - 1992/7/24

Y1 - 1992/7/24

N2 - An expression vector was modified to permit the rapid synthesis of purified, 32P-labeled, glutathione S-transferase (GST)-retinoblastoma (RB) fusion proteins. The products were used to screen λgt11 expression libraries, from which we cloned a cDNA encoding a polypeptide (RBAP-1) capable of binding directly to a putative functional domain (the pocket) of the retinoblastoma gene product (RB). The RB "pocket" is known to bind, directly or indirectly, to the cellular transcription factor, E2F, implicated in cell growth control. We have found that RBAP-1 copurifies with E2F, interacts specifically with the adenovirus E4 ORF 6 7 protein, binds specifically and directly to a known E2F DNA recognition sequence, and contains a functional transactivation domain. Therefore, RBAP-1 is a species of E2F and can bind specifically to the RB pocket.

AB - An expression vector was modified to permit the rapid synthesis of purified, 32P-labeled, glutathione S-transferase (GST)-retinoblastoma (RB) fusion proteins. The products were used to screen λgt11 expression libraries, from which we cloned a cDNA encoding a polypeptide (RBAP-1) capable of binding directly to a putative functional domain (the pocket) of the retinoblastoma gene product (RB). The RB "pocket" is known to bind, directly or indirectly, to the cellular transcription factor, E2F, implicated in cell growth control. We have found that RBAP-1 copurifies with E2F, interacts specifically with the adenovirus E4 ORF 6 7 protein, binds specifically and directly to a known E2F DNA recognition sequence, and contains a functional transactivation domain. Therefore, RBAP-1 is a species of E2F and can bind specifically to the RB pocket.

UR - http://www.scopus.com/inward/record.url?scp=0026766083&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026766083&partnerID=8YFLogxK

U2 - 10.1016/0092-8674(92)90108-O

DO - 10.1016/0092-8674(92)90108-O

M3 - Article

C2 - 1638635

AN - SCOPUS:0026766083

VL - 70

SP - 351

EP - 364

JO - Cell

JF - Cell

SN - 0092-8674

IS - 2

ER -