Wild-type human p53 and a temperature-sensitive mutant induce Fas/APO-1 expression

L. B. Owen-Schaub, W. Zhang, J. C. Cusack, L. S. Angelo, S. M. Santee, Toshiyoshi Fujiwara, J. A. Roth, A. B. Deisseroth, W. W. Zhang, E. Kruzel, R. Radinsky

Research output: Contribution to journalArticle

670 Citations (Scopus)

Abstract

Fas/APO-1 is a cell surface protein known to trigger apoptosis upon specific antibody engagement. Because wild-type p53 can activate transcription as well as induce apoptosis, we queried whether p53 might upregulate Fas/APO-1. To explore this possibility, we examined human p53- null (H358 non-small-cell lung adenocarcinoma and K562 erythroleukemia) and wild-type p53-containing (H460 non-small-cell lung adenocarcinoma) cell lines. When H358 or H460 cells were transduced with a replication-deficient adenovirus expression construct containing the human wild-type p53 gene but not with vector alone, a marked upregulation (approximately a three- to fourfold increase) of cell surface Fas/APO-1 was observed by flow cytometry. Similarly, K562 cells stably transfected with a plasmid vector containing the temperature-sensitive human p53 mutant Ala-143 demonstrated a four- to sixfold upregulation of Fas/APO-1 by flow-cytometric analysis at the permissive temperature of 32.5°C. Temperature-sensitive upregulation of Fas/APO-1 in K562 Ala-143 cells was verified by immunoprecipitation and demonstrated to result from enhanced mRNA production by nuclear run-on and Northern (RNA) analyses. Stably transfected K562 cells expressing temperature-insensitive, transcriptionally inactive p53 mutants (His-175, Trp-248, His-273, or Gly-281) failed to upregulate Fas/APO-1 at either 32.5° or 37.5°C. The temperature-sensitive transcription of Fas/APO-1 occurred in the presence of cycloheximide, indicating that de novo protein synthesis was not required and suggested a direct involvement of p53. Collectively, these observations argue that Fas/APO-1 is a target gene for transcriptional activation by p53.

Original languageEnglish
Pages (from-to)3032-3040
Number of pages9
JournalMolecular and Cellular Biology
Volume15
Issue number6
Publication statusPublished - 1995
Externally publishedYes

Fingerprint

Up-Regulation
Temperature
K562 Cells
Transcriptional Activation
Apoptosis
Leukemia, Erythroblastic, Acute
p53 Genes
Cycloheximide
Immunoprecipitation
Adenoviridae
Flow Cytometry
Membrane Proteins
Plasmids
RNA
Cell Line
Messenger RNA
Antibodies
Proteins
Adenocarcinoma of lung

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Owen-Schaub, L. B., Zhang, W., Cusack, J. C., Angelo, L. S., Santee, S. M., Fujiwara, T., ... Radinsky, R. (1995). Wild-type human p53 and a temperature-sensitive mutant induce Fas/APO-1 expression. Molecular and Cellular Biology, 15(6), 3032-3040.

Wild-type human p53 and a temperature-sensitive mutant induce Fas/APO-1 expression. / Owen-Schaub, L. B.; Zhang, W.; Cusack, J. C.; Angelo, L. S.; Santee, S. M.; Fujiwara, Toshiyoshi; Roth, J. A.; Deisseroth, A. B.; Zhang, W. W.; Kruzel, E.; Radinsky, R.

In: Molecular and Cellular Biology, Vol. 15, No. 6, 1995, p. 3032-3040.

Research output: Contribution to journalArticle

Owen-Schaub, LB, Zhang, W, Cusack, JC, Angelo, LS, Santee, SM, Fujiwara, T, Roth, JA, Deisseroth, AB, Zhang, WW, Kruzel, E & Radinsky, R 1995, 'Wild-type human p53 and a temperature-sensitive mutant induce Fas/APO-1 expression', Molecular and Cellular Biology, vol. 15, no. 6, pp. 3032-3040.
Owen-Schaub LB, Zhang W, Cusack JC, Angelo LS, Santee SM, Fujiwara T et al. Wild-type human p53 and a temperature-sensitive mutant induce Fas/APO-1 expression. Molecular and Cellular Biology. 1995;15(6):3032-3040.
Owen-Schaub, L. B. ; Zhang, W. ; Cusack, J. C. ; Angelo, L. S. ; Santee, S. M. ; Fujiwara, Toshiyoshi ; Roth, J. A. ; Deisseroth, A. B. ; Zhang, W. W. ; Kruzel, E. ; Radinsky, R. / Wild-type human p53 and a temperature-sensitive mutant induce Fas/APO-1 expression. In: Molecular and Cellular Biology. 1995 ; Vol. 15, No. 6. pp. 3032-3040.
@article{7737406bfc4e45eba24d607010d07504,
title = "Wild-type human p53 and a temperature-sensitive mutant induce Fas/APO-1 expression",
abstract = "Fas/APO-1 is a cell surface protein known to trigger apoptosis upon specific antibody engagement. Because wild-type p53 can activate transcription as well as induce apoptosis, we queried whether p53 might upregulate Fas/APO-1. To explore this possibility, we examined human p53- null (H358 non-small-cell lung adenocarcinoma and K562 erythroleukemia) and wild-type p53-containing (H460 non-small-cell lung adenocarcinoma) cell lines. When H358 or H460 cells were transduced with a replication-deficient adenovirus expression construct containing the human wild-type p53 gene but not with vector alone, a marked upregulation (approximately a three- to fourfold increase) of cell surface Fas/APO-1 was observed by flow cytometry. Similarly, K562 cells stably transfected with a plasmid vector containing the temperature-sensitive human p53 mutant Ala-143 demonstrated a four- to sixfold upregulation of Fas/APO-1 by flow-cytometric analysis at the permissive temperature of 32.5°C. Temperature-sensitive upregulation of Fas/APO-1 in K562 Ala-143 cells was verified by immunoprecipitation and demonstrated to result from enhanced mRNA production by nuclear run-on and Northern (RNA) analyses. Stably transfected K562 cells expressing temperature-insensitive, transcriptionally inactive p53 mutants (His-175, Trp-248, His-273, or Gly-281) failed to upregulate Fas/APO-1 at either 32.5° or 37.5°C. The temperature-sensitive transcription of Fas/APO-1 occurred in the presence of cycloheximide, indicating that de novo protein synthesis was not required and suggested a direct involvement of p53. Collectively, these observations argue that Fas/APO-1 is a target gene for transcriptional activation by p53.",
author = "Owen-Schaub, {L. B.} and W. Zhang and Cusack, {J. C.} and Angelo, {L. S.} and Santee, {S. M.} and Toshiyoshi Fujiwara and Roth, {J. A.} and Deisseroth, {A. B.} and Zhang, {W. W.} and E. Kruzel and R. Radinsky",
year = "1995",
language = "English",
volume = "15",
pages = "3032--3040",
journal = "Molecular and Cellular Biology",
issn = "0270-7306",
publisher = "American Society for Microbiology",
number = "6",

}

TY - JOUR

T1 - Wild-type human p53 and a temperature-sensitive mutant induce Fas/APO-1 expression

AU - Owen-Schaub, L. B.

AU - Zhang, W.

AU - Cusack, J. C.

AU - Angelo, L. S.

AU - Santee, S. M.

AU - Fujiwara, Toshiyoshi

AU - Roth, J. A.

AU - Deisseroth, A. B.

AU - Zhang, W. W.

AU - Kruzel, E.

AU - Radinsky, R.

PY - 1995

Y1 - 1995

N2 - Fas/APO-1 is a cell surface protein known to trigger apoptosis upon specific antibody engagement. Because wild-type p53 can activate transcription as well as induce apoptosis, we queried whether p53 might upregulate Fas/APO-1. To explore this possibility, we examined human p53- null (H358 non-small-cell lung adenocarcinoma and K562 erythroleukemia) and wild-type p53-containing (H460 non-small-cell lung adenocarcinoma) cell lines. When H358 or H460 cells were transduced with a replication-deficient adenovirus expression construct containing the human wild-type p53 gene but not with vector alone, a marked upregulation (approximately a three- to fourfold increase) of cell surface Fas/APO-1 was observed by flow cytometry. Similarly, K562 cells stably transfected with a plasmid vector containing the temperature-sensitive human p53 mutant Ala-143 demonstrated a four- to sixfold upregulation of Fas/APO-1 by flow-cytometric analysis at the permissive temperature of 32.5°C. Temperature-sensitive upregulation of Fas/APO-1 in K562 Ala-143 cells was verified by immunoprecipitation and demonstrated to result from enhanced mRNA production by nuclear run-on and Northern (RNA) analyses. Stably transfected K562 cells expressing temperature-insensitive, transcriptionally inactive p53 mutants (His-175, Trp-248, His-273, or Gly-281) failed to upregulate Fas/APO-1 at either 32.5° or 37.5°C. The temperature-sensitive transcription of Fas/APO-1 occurred in the presence of cycloheximide, indicating that de novo protein synthesis was not required and suggested a direct involvement of p53. Collectively, these observations argue that Fas/APO-1 is a target gene for transcriptional activation by p53.

AB - Fas/APO-1 is a cell surface protein known to trigger apoptosis upon specific antibody engagement. Because wild-type p53 can activate transcription as well as induce apoptosis, we queried whether p53 might upregulate Fas/APO-1. To explore this possibility, we examined human p53- null (H358 non-small-cell lung adenocarcinoma and K562 erythroleukemia) and wild-type p53-containing (H460 non-small-cell lung adenocarcinoma) cell lines. When H358 or H460 cells were transduced with a replication-deficient adenovirus expression construct containing the human wild-type p53 gene but not with vector alone, a marked upregulation (approximately a three- to fourfold increase) of cell surface Fas/APO-1 was observed by flow cytometry. Similarly, K562 cells stably transfected with a plasmid vector containing the temperature-sensitive human p53 mutant Ala-143 demonstrated a four- to sixfold upregulation of Fas/APO-1 by flow-cytometric analysis at the permissive temperature of 32.5°C. Temperature-sensitive upregulation of Fas/APO-1 in K562 Ala-143 cells was verified by immunoprecipitation and demonstrated to result from enhanced mRNA production by nuclear run-on and Northern (RNA) analyses. Stably transfected K562 cells expressing temperature-insensitive, transcriptionally inactive p53 mutants (His-175, Trp-248, His-273, or Gly-281) failed to upregulate Fas/APO-1 at either 32.5° or 37.5°C. The temperature-sensitive transcription of Fas/APO-1 occurred in the presence of cycloheximide, indicating that de novo protein synthesis was not required and suggested a direct involvement of p53. Collectively, these observations argue that Fas/APO-1 is a target gene for transcriptional activation by p53.

UR - http://www.scopus.com/inward/record.url?scp=0029003501&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029003501&partnerID=8YFLogxK

M3 - Article

C2 - 7539102

AN - SCOPUS:0029003501

VL - 15

SP - 3032

EP - 3040

JO - Molecular and Cellular Biology

JF - Molecular and Cellular Biology

SN - 0270-7306

IS - 6

ER -