P300 regulates the human RLIP76 promoter activity and gene expression

Archana Sehrawat, Sushma Yadav, Yogesh C. Awasthi, Alakananda Basu, Charles Warden, Sanjay Awasthi

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

A 76-kDa Ral-interacting protein (RLIP76) has been implicated in the pathogenesis of cancer and diabetes. It is often over expressed in human malignant cell lines and human tumor samples and has been associated with metastasis and chemoresistance. RLIP76 homozygous knockout mice exhibit increased insulin sensitivity, hypoglycemia, and hypolipidemia, and resist cancer development. Little is known about the mechanism by which the expression of RLIP76 is regulated. In the present study, we functionally characterized the RLIP76 promoter using deletion mapping and mutational analysis to investigate the regulation of RLIP76 transcription. We have identified the promoter regions important for RLIP76 transcription, including a strong cis-activating element in the proximal promoter containing overlapping consensus cMYB and cETS binding sites. Transcription factor cMYB and the coactivator p300 associated with RLIP76 gene promoter as shown by CHIP assay. Knockdown of p300 in HEK293 cells reduced the activity of the promoter fragment containing wild type cMYB/cETS binding site in comparison to that with deleted or mutated cMYB/cETS binding site. Knockdown of p300 also decreased the RLIP76 expression as indicated by immunoblotting, immunocytochemistry and flow cytometry analysis. Thus, we report for the first time that p300 associates with the RLIP76 promoter via an overlapping cMYB and cETS binding site and regulates RLIP76 promoter activity and its expression.

Original languageEnglish
Pages (from-to)1203-1211
Number of pages9
JournalBiochemical Pharmacology
Volume85
Issue number8
DOIs
StatePublished - 15 Apr 2013

Fingerprint

Gene expression
Binding Sites
Gene Expression
Transcription
ral GTP-Binding Proteins
Flow cytometry
HEK293 Cells
Medical problems
Tumor Cell Line
Hypoglycemia
Immunoblotting
Genetic Promoter Regions
Knockout Mice
Insulin Resistance
Tumors
Assays
Neoplasms
Flow Cytometry
Transcription Factors
Genes

Keywords

  • Promoter activity
  • RLIP76
  • Transcription
  • cETS
  • cMYB
  • p300

Cite this

Sehrawat, A., Yadav, S., Awasthi, Y. C., Basu, A., Warden, C., & Awasthi, S. (2013). P300 regulates the human RLIP76 promoter activity and gene expression. Biochemical Pharmacology, 85(8), 1203-1211. https://doi.org/10.1016/j.bcp.2013.02.012
Sehrawat, Archana ; Yadav, Sushma ; Awasthi, Yogesh C. ; Basu, Alakananda ; Warden, Charles ; Awasthi, Sanjay. / P300 regulates the human RLIP76 promoter activity and gene expression. In: Biochemical Pharmacology. 2013 ; Vol. 85, No. 8. pp. 1203-1211.
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Sehrawat, A, Yadav, S, Awasthi, YC, Basu, A, Warden, C & Awasthi, S 2013, 'P300 regulates the human RLIP76 promoter activity and gene expression', Biochemical Pharmacology, vol. 85, no. 8, pp. 1203-1211. https://doi.org/10.1016/j.bcp.2013.02.012

P300 regulates the human RLIP76 promoter activity and gene expression. / Sehrawat, Archana; Yadav, Sushma; Awasthi, Yogesh C.; Basu, Alakananda; Warden, Charles; Awasthi, Sanjay.

In: Biochemical Pharmacology, Vol. 85, No. 8, 15.04.2013, p. 1203-1211.

Research output: Contribution to journalArticle

TY - JOUR

T1 - P300 regulates the human RLIP76 promoter activity and gene expression

AU - Sehrawat, Archana

AU - Yadav, Sushma

AU - Awasthi, Yogesh C.

AU - Basu, Alakananda

AU - Warden, Charles

AU - Awasthi, Sanjay

PY - 2013/4/15

Y1 - 2013/4/15

N2 - A 76-kDa Ral-interacting protein (RLIP76) has been implicated in the pathogenesis of cancer and diabetes. It is often over expressed in human malignant cell lines and human tumor samples and has been associated with metastasis and chemoresistance. RLIP76 homozygous knockout mice exhibit increased insulin sensitivity, hypoglycemia, and hypolipidemia, and resist cancer development. Little is known about the mechanism by which the expression of RLIP76 is regulated. In the present study, we functionally characterized the RLIP76 promoter using deletion mapping and mutational analysis to investigate the regulation of RLIP76 transcription. We have identified the promoter regions important for RLIP76 transcription, including a strong cis-activating element in the proximal promoter containing overlapping consensus cMYB and cETS binding sites. Transcription factor cMYB and the coactivator p300 associated with RLIP76 gene promoter as shown by CHIP assay. Knockdown of p300 in HEK293 cells reduced the activity of the promoter fragment containing wild type cMYB/cETS binding site in comparison to that with deleted or mutated cMYB/cETS binding site. Knockdown of p300 also decreased the RLIP76 expression as indicated by immunoblotting, immunocytochemistry and flow cytometry analysis. Thus, we report for the first time that p300 associates with the RLIP76 promoter via an overlapping cMYB and cETS binding site and regulates RLIP76 promoter activity and its expression.

AB - A 76-kDa Ral-interacting protein (RLIP76) has been implicated in the pathogenesis of cancer and diabetes. It is often over expressed in human malignant cell lines and human tumor samples and has been associated with metastasis and chemoresistance. RLIP76 homozygous knockout mice exhibit increased insulin sensitivity, hypoglycemia, and hypolipidemia, and resist cancer development. Little is known about the mechanism by which the expression of RLIP76 is regulated. In the present study, we functionally characterized the RLIP76 promoter using deletion mapping and mutational analysis to investigate the regulation of RLIP76 transcription. We have identified the promoter regions important for RLIP76 transcription, including a strong cis-activating element in the proximal promoter containing overlapping consensus cMYB and cETS binding sites. Transcription factor cMYB and the coactivator p300 associated with RLIP76 gene promoter as shown by CHIP assay. Knockdown of p300 in HEK293 cells reduced the activity of the promoter fragment containing wild type cMYB/cETS binding site in comparison to that with deleted or mutated cMYB/cETS binding site. Knockdown of p300 also decreased the RLIP76 expression as indicated by immunoblotting, immunocytochemistry and flow cytometry analysis. Thus, we report for the first time that p300 associates with the RLIP76 promoter via an overlapping cMYB and cETS binding site and regulates RLIP76 promoter activity and its expression.

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KW - RLIP76

KW - Transcription

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KW - cMYB

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DO - 10.1016/j.bcp.2013.02.012

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