Functional expression and analysis of the pancreatic transcription factor PDX-1 in yeast

Sabire Ozcan; Amber L. Mosley; Bishwa K. Aryal

doi:10.1016/S0006-291X(02)00747-7

Functional expression and analysis of the pancreatic transcription factor PDX-1 in yeast

Sabire Ozcan, Amber L. Mosley, Bishwa K. Aryal

Molecular and Cellular Biochemistry

Producción científica: Article › revisión exhaustiva

2 Citas (Scopus)

Resumen

The pancreas-specific transcription factor Pdx-1 is important for pancreas development and β-cell specific gene expression in insulin-producing cells. We have expressed the mouse PDX-1 gene in the yeast Saccharomyces cerevisiae and characterized its functional domains. Pdx-1 functions as a strong activator in yeast and stimulates gene expression by more than 80-fold. The transcriptional activation domain of Pdx-1 is located within the first 144 amino-terminal amino acids. Pdx-1 is also able to bind and activate transcription from the A3 element of the human insulin gene promoter in yeast. Analysis of the effects of two-point mutations (Q59L and R197H) in the PDX-1 gene found in type II diabetes patients showed that both point mutations interfere with the ability of Pdx-1 to bind to DNA and to activate transcription in yeast.

Idioma original	English
Páginas (desde-hasta)	724-729
Número de páginas	6
Publicación	Biochemical and Biophysical Research Communications
Volumen	295
N.º	3
DOI	https://doi.org/10.1016/S0006-291X(02)00747-7
Estado	Published - 2002

Nota bibliográfica

Funding Information:
This work was supported by a Juvenile Diabetes Foundation Career Development Award (to S.O.) We thank Alan Permutt (Washington University, St. Louis) for providing the cDNA for the mouse PDX-1 gene, Michael German (UCSD) for the plasmid containing the A3 region, and Fang Fang for excellent technical assistance.

Financiación

This work was supported by a Juvenile Diabetes Foundation Career Development Award (to S.O.) We thank Alan Permutt (Washington University, St. Louis) for providing the cDNA for the mouse PDX-1 gene, Michael German (UCSD) for the plasmid containing the A3 region, and Fang Fang for excellent technical assistance.

Financiadores	Número del financiador
University of California San Diego Health
Juvenile Diabetes Research Foundation United States of America

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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title = "Functional expression and analysis of the pancreatic transcription factor PDX-1 in yeast",

abstract = "The pancreas-specific transcription factor Pdx-1 is important for pancreas development and β-cell specific gene expression in insulin-producing cells. We have expressed the mouse PDX-1 gene in the yeast Saccharomyces cerevisiae and characterized its functional domains. Pdx-1 functions as a strong activator in yeast and stimulates gene expression by more than 80-fold. The transcriptional activation domain of Pdx-1 is located within the first 144 amino-terminal amino acids. Pdx-1 is also able to bind and activate transcription from the A3 element of the human insulin gene promoter in yeast. Analysis of the effects of two-point mutations (Q59L and R197H) in the PDX-1 gene found in type II diabetes patients showed that both point mutations interfere with the ability of Pdx-1 to bind to DNA and to activate transcription in yeast.",

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author = "Sabire Ozcan and Mosley, \{Amber L.\} and Aryal, \{Bishwa K.\}",

note = "Funding Information: This work was supported by a Juvenile Diabetes Foundation Career Development Award (to S.O.) We thank Alan Permutt (Washington University, St. Louis) for providing the cDNA for the mouse PDX-1 gene, Michael German (UCSD) for the plasmid containing the A3 region, and Fang Fang for excellent technical assistance.",

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AU - Mosley, Amber L.

AU - Aryal, Bishwa K.

N1 - Funding Information: This work was supported by a Juvenile Diabetes Foundation Career Development Award (to S.O.) We thank Alan Permutt (Washington University, St. Louis) for providing the cDNA for the mouse PDX-1 gene, Michael German (UCSD) for the plasmid containing the A3 region, and Fang Fang for excellent technical assistance.

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N2 - The pancreas-specific transcription factor Pdx-1 is important for pancreas development and β-cell specific gene expression in insulin-producing cells. We have expressed the mouse PDX-1 gene in the yeast Saccharomyces cerevisiae and characterized its functional domains. Pdx-1 functions as a strong activator in yeast and stimulates gene expression by more than 80-fold. The transcriptional activation domain of Pdx-1 is located within the first 144 amino-terminal amino acids. Pdx-1 is also able to bind and activate transcription from the A3 element of the human insulin gene promoter in yeast. Analysis of the effects of two-point mutations (Q59L and R197H) in the PDX-1 gene found in type II diabetes patients showed that both point mutations interfere with the ability of Pdx-1 to bind to DNA and to activate transcription in yeast.

AB - The pancreas-specific transcription factor Pdx-1 is important for pancreas development and β-cell specific gene expression in insulin-producing cells. We have expressed the mouse PDX-1 gene in the yeast Saccharomyces cerevisiae and characterized its functional domains. Pdx-1 functions as a strong activator in yeast and stimulates gene expression by more than 80-fold. The transcriptional activation domain of Pdx-1 is located within the first 144 amino-terminal amino acids. Pdx-1 is also able to bind and activate transcription from the A3 element of the human insulin gene promoter in yeast. Analysis of the effects of two-point mutations (Q59L and R197H) in the PDX-1 gene found in type II diabetes patients showed that both point mutations interfere with the ability of Pdx-1 to bind to DNA and to activate transcription in yeast.

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KW - Gene expression

KW - Pancreas

KW - Pdx-1

KW - Transcription factor

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Functional expression and analysis of the pancreatic transcription factor PDX-1 in yeast

Resumen

Nota bibliográfica

Financiación

ASJC Scopus subject areas

ODS de las Naciones Unidas

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