Functional comparison of the Mus musculus molossinus and Mus musculus domesticus Sry genes

R. A. Dubin, P. Coward, Y. F C Lau, Harry Ostrer

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The Sry gene functions as a genetic switch initiating testicular development of the indifferent mammalian gonad. The Bus musculus molossinus Sry open reading frame (ORF) encodes a 395-amino acid transcription factor (mSry) that specifically binds and bends DNA through its N-terminal HMG domain and activates transcription through its long C-terminal (residues 144- 366) glutamine/histidine-rich activation domain. The M. m. domesticus Sty ORF encodes a highly homologous, truncated protein (dSry) of approximately 230 amino acids, and the molecular basis for truncation is a point mutation that creates an amber stop codon within the activation domain. The mSry protein activates transcription of a Sry-responsive reporter gene in HeLa cells, but dSry does not. Gene swapping and in vitro DNA binding experiments revealed that lack of transcriptional activation by dSry was not the result of polymorphisms within the first 137 amino acids of the protein. Direct analysis of the C-terminal glutamine/histidine-rich domain revealed that dSry lacked a functional transcriptional activation domain. Fusion of the GAL4 DNA-binding domain to the C-terminal region of mSry created a potent transactivator. By contrast, a similar GAL4-dSry protein exhibited severely reduced activity. Analysis of a series of C-terminal deletion mutants of the GAL4-mSry chimeric protein indicated that residues 263-345 of the glutamine/histidine-rich domain were necessary for high level transactivation. Furthermore, read through of the premature amber stop codon by transfer RNA suppression resulted in a strong GAL4-dSry transactivator. This demonstrated that the premature stop codon is the only polymorphism responsible for the inability of the dSry glutamine/histidine-rich region to transactivate.

Original languageEnglish (US)
Pages (from-to)1645-1654
Number of pages10
JournalMolecular Endocrinology
Volume9
Issue number12
StatePublished - 1995
Externally publishedYes

Fingerprint

sry Genes
Glutamine
Histidine
Transcriptional Activation
Trans-Activators
Terminator Codon
Nonsense Codon
Proteins
Amino Acids
Open Reading Frames
Hordeolum
DNA
Gonads
Motor Vehicles
Transfer RNA
Reporter Genes
HeLa Cells
Point Mutation
Transcription Factors
Genes

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Functional comparison of the Mus musculus molossinus and Mus musculus domesticus Sry genes. / Dubin, R. A.; Coward, P.; Lau, Y. F C; Ostrer, Harry.

In: Molecular Endocrinology, Vol. 9, No. 12, 1995, p. 1645-1654.

Research output: Contribution to journalArticle

Dubin, R. A. ; Coward, P. ; Lau, Y. F C ; Ostrer, Harry. / Functional comparison of the Mus musculus molossinus and Mus musculus domesticus Sry genes. In: Molecular Endocrinology. 1995 ; Vol. 9, No. 12. pp. 1645-1654.
@article{e6ad2ca7039e454a8c027663155a2756,
title = "Functional comparison of the Mus musculus molossinus and Mus musculus domesticus Sry genes",
abstract = "The Sry gene functions as a genetic switch initiating testicular development of the indifferent mammalian gonad. The Bus musculus molossinus Sry open reading frame (ORF) encodes a 395-amino acid transcription factor (mSry) that specifically binds and bends DNA through its N-terminal HMG domain and activates transcription through its long C-terminal (residues 144- 366) glutamine/histidine-rich activation domain. The M. m. domesticus Sty ORF encodes a highly homologous, truncated protein (dSry) of approximately 230 amino acids, and the molecular basis for truncation is a point mutation that creates an amber stop codon within the activation domain. The mSry protein activates transcription of a Sry-responsive reporter gene in HeLa cells, but dSry does not. Gene swapping and in vitro DNA binding experiments revealed that lack of transcriptional activation by dSry was not the result of polymorphisms within the first 137 amino acids of the protein. Direct analysis of the C-terminal glutamine/histidine-rich domain revealed that dSry lacked a functional transcriptional activation domain. Fusion of the GAL4 DNA-binding domain to the C-terminal region of mSry created a potent transactivator. By contrast, a similar GAL4-dSry protein exhibited severely reduced activity. Analysis of a series of C-terminal deletion mutants of the GAL4-mSry chimeric protein indicated that residues 263-345 of the glutamine/histidine-rich domain were necessary for high level transactivation. Furthermore, read through of the premature amber stop codon by transfer RNA suppression resulted in a strong GAL4-dSry transactivator. This demonstrated that the premature stop codon is the only polymorphism responsible for the inability of the dSry glutamine/histidine-rich region to transactivate.",
author = "Dubin, {R. A.} and P. Coward and Lau, {Y. F C} and Harry Ostrer",
year = "1995",
language = "English (US)",
volume = "9",
pages = "1645--1654",
journal = "Molecular Endocrinology",
issn = "0888-8809",
publisher = "The Endocrine Society",
number = "12",

}

TY - JOUR

T1 - Functional comparison of the Mus musculus molossinus and Mus musculus domesticus Sry genes

AU - Dubin, R. A.

AU - Coward, P.

AU - Lau, Y. F C

AU - Ostrer, Harry

PY - 1995

Y1 - 1995

N2 - The Sry gene functions as a genetic switch initiating testicular development of the indifferent mammalian gonad. The Bus musculus molossinus Sry open reading frame (ORF) encodes a 395-amino acid transcription factor (mSry) that specifically binds and bends DNA through its N-terminal HMG domain and activates transcription through its long C-terminal (residues 144- 366) glutamine/histidine-rich activation domain. The M. m. domesticus Sty ORF encodes a highly homologous, truncated protein (dSry) of approximately 230 amino acids, and the molecular basis for truncation is a point mutation that creates an amber stop codon within the activation domain. The mSry protein activates transcription of a Sry-responsive reporter gene in HeLa cells, but dSry does not. Gene swapping and in vitro DNA binding experiments revealed that lack of transcriptional activation by dSry was not the result of polymorphisms within the first 137 amino acids of the protein. Direct analysis of the C-terminal glutamine/histidine-rich domain revealed that dSry lacked a functional transcriptional activation domain. Fusion of the GAL4 DNA-binding domain to the C-terminal region of mSry created a potent transactivator. By contrast, a similar GAL4-dSry protein exhibited severely reduced activity. Analysis of a series of C-terminal deletion mutants of the GAL4-mSry chimeric protein indicated that residues 263-345 of the glutamine/histidine-rich domain were necessary for high level transactivation. Furthermore, read through of the premature amber stop codon by transfer RNA suppression resulted in a strong GAL4-dSry transactivator. This demonstrated that the premature stop codon is the only polymorphism responsible for the inability of the dSry glutamine/histidine-rich region to transactivate.

AB - The Sry gene functions as a genetic switch initiating testicular development of the indifferent mammalian gonad. The Bus musculus molossinus Sry open reading frame (ORF) encodes a 395-amino acid transcription factor (mSry) that specifically binds and bends DNA through its N-terminal HMG domain and activates transcription through its long C-terminal (residues 144- 366) glutamine/histidine-rich activation domain. The M. m. domesticus Sty ORF encodes a highly homologous, truncated protein (dSry) of approximately 230 amino acids, and the molecular basis for truncation is a point mutation that creates an amber stop codon within the activation domain. The mSry protein activates transcription of a Sry-responsive reporter gene in HeLa cells, but dSry does not. Gene swapping and in vitro DNA binding experiments revealed that lack of transcriptional activation by dSry was not the result of polymorphisms within the first 137 amino acids of the protein. Direct analysis of the C-terminal glutamine/histidine-rich domain revealed that dSry lacked a functional transcriptional activation domain. Fusion of the GAL4 DNA-binding domain to the C-terminal region of mSry created a potent transactivator. By contrast, a similar GAL4-dSry protein exhibited severely reduced activity. Analysis of a series of C-terminal deletion mutants of the GAL4-mSry chimeric protein indicated that residues 263-345 of the glutamine/histidine-rich domain were necessary for high level transactivation. Furthermore, read through of the premature amber stop codon by transfer RNA suppression resulted in a strong GAL4-dSry transactivator. This demonstrated that the premature stop codon is the only polymorphism responsible for the inability of the dSry glutamine/histidine-rich region to transactivate.

UR - http://www.scopus.com/inward/record.url?scp=0028842893&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028842893&partnerID=8YFLogxK

M3 - Article

C2 - 8614401

AN - SCOPUS:0028842893

VL - 9

SP - 1645

EP - 1654

JO - Molecular Endocrinology

JF - Molecular Endocrinology

SN - 0888-8809

IS - 12

ER -