Functional redundancy of octamer elements in the mouse mammary tumor virus promoter

Mike Huang; Jae Woon Lee; David O. Peterson

doi:10.1093/nar/21.22.5235

Functional redundancy of octamer elements in the mouse mammary tumor virus promoter

Mike Huang, Jae Woon Lee, David O. Peterson

Research output: Contribution to journal › Article › peer-review

12 Scopus citations

Abstract

The promoter of mouse mammary tumor virus contains three overlapping sequence elements related to the octamer consensus (ATGCAAAT) that are largely contained within two 10 bp direct repeats (CTTATGTAAA) separated by a 2 bp spacer between 60 and 39 relative to the start of transcription. Gel electrophoresis mobility shift competition assays demonstrate that the most distal of these octamer-related elements is recognized by a protein that also binds to the most proximal element, while the central octamer-related element is not efficiently recognized. Transient transfection assays with altered promoters reveal that the portion of the 10 bp repeat that is not related to the octamer consensus appears not to be important for transcription. The distal and proximal octamer-related elements are, at least to some extent, functionally redundant. Complete deletion of one element has little or no effect on promoter activity so long as certain spacing constraints among remaining promoter elements are maintained. Systematic variation of such spacing reveals a cyclic effect on promoter activity corresponding to the periodicity of Bform DNA, suggesting functional interactions between proteins bound to adjacent sites.

Original language	English (US)
Pages (from-to)	5235-5241
Number of pages	7
Journal	Nucleic acids research
Volume	21
Issue number	22
DOIs	https://doi.org/10.1093/nar/21.22.5235
State	Published - Nov 11 1993
Externally published	Yes

ASJC Scopus subject areas

Genetics

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title = "Functional redundancy of octamer elements in the mouse mammary tumor virus promoter",

abstract = "The promoter of mouse mammary tumor virus contains three overlapping sequence elements related to the octamer consensus (ATGCAAAT) that are largely contained within two 10 bp direct repeats (CTTATGTAAA) separated by a 2 bp spacer between 60 and 39 relative to the start of transcription. Gel electrophoresis mobility shift competition assays demonstrate that the most distal of these octamer-related elements is recognized by a protein that also binds to the most proximal element, while the central octamer-related element is not efficiently recognized. Transient transfection assays with altered promoters reveal that the portion of the 10 bp repeat that is not related to the octamer consensus appears not to be important for transcription. The distal and proximal octamer-related elements are, at least to some extent, functionally redundant. Complete deletion of one element has little or no effect on promoter activity so long as certain spacing constraints among remaining promoter elements are maintained. Systematic variation of such spacing reveals a cyclic effect on promoter activity corresponding to the periodicity of Bform DNA, suggesting functional interactions between proteins bound to adjacent sites.",

author = "Mike Huang and Lee, {Jae Woon} and Peterson, {David O.}",

note = "Funding Information: We thank Katherine Beifuss for expert technical assistance and Gary Kunkel for helpful comments on the manuscript. This work was supported by Public Health Service Grants CA32695 and CA48041 from the National Cancer Institute. The support of the Texas Agricultural Experiment Station is also gratefully acknowledged.",

year = "1993",

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doi = "10.1093/nar/21.22.5235",

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AU - Huang, Mike

AU - Lee, Jae Woon

AU - Peterson, David O.

N1 - Funding Information: We thank Katherine Beifuss for expert technical assistance and Gary Kunkel for helpful comments on the manuscript. This work was supported by Public Health Service Grants CA32695 and CA48041 from the National Cancer Institute. The support of the Texas Agricultural Experiment Station is also gratefully acknowledged.

PY - 1993/11/11

Y1 - 1993/11/11

N2 - The promoter of mouse mammary tumor virus contains three overlapping sequence elements related to the octamer consensus (ATGCAAAT) that are largely contained within two 10 bp direct repeats (CTTATGTAAA) separated by a 2 bp spacer between 60 and 39 relative to the start of transcription. Gel electrophoresis mobility shift competition assays demonstrate that the most distal of these octamer-related elements is recognized by a protein that also binds to the most proximal element, while the central octamer-related element is not efficiently recognized. Transient transfection assays with altered promoters reveal that the portion of the 10 bp repeat that is not related to the octamer consensus appears not to be important for transcription. The distal and proximal octamer-related elements are, at least to some extent, functionally redundant. Complete deletion of one element has little or no effect on promoter activity so long as certain spacing constraints among remaining promoter elements are maintained. Systematic variation of such spacing reveals a cyclic effect on promoter activity corresponding to the periodicity of Bform DNA, suggesting functional interactions between proteins bound to adjacent sites.

AB - The promoter of mouse mammary tumor virus contains three overlapping sequence elements related to the octamer consensus (ATGCAAAT) that are largely contained within two 10 bp direct repeats (CTTATGTAAA) separated by a 2 bp spacer between 60 and 39 relative to the start of transcription. Gel electrophoresis mobility shift competition assays demonstrate that the most distal of these octamer-related elements is recognized by a protein that also binds to the most proximal element, while the central octamer-related element is not efficiently recognized. Transient transfection assays with altered promoters reveal that the portion of the 10 bp repeat that is not related to the octamer consensus appears not to be important for transcription. The distal and proximal octamer-related elements are, at least to some extent, functionally redundant. Complete deletion of one element has little or no effect on promoter activity so long as certain spacing constraints among remaining promoter elements are maintained. Systematic variation of such spacing reveals a cyclic effect on promoter activity corresponding to the periodicity of Bform DNA, suggesting functional interactions between proteins bound to adjacent sites.

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Functional redundancy of octamer elements in the mouse mammary tumor virus promoter

Abstract

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