Identification of a negative regulatory element that inhibits c-mos transcription in somatic cells

Sandra S. Zinkel, Subrata K. Pal, J. Szeberényi, Geoffrey M. Cooper

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

We have used transient expression assays to identify a cis-acting region in the 5′ flanking sequence of murine c-mos which, when deleted, allows expression from the c-mos promoter in NIH 3T3 cells. This negative regulatory sequence, located 400 to 500 nucleotides upstream of the c-mos ATG, also inhibited expression from a heterologous promoter. In addition to NIH 3T3 cells, the c-mos negative regulatory sequence was active in BALB/3T3 cells, PC12 rat pheochromocytoma cells, and A549 human lung carcinoma cells. Site-specific mutagenesis identified three possibly interacting regions that were involved in negative regulatory activity, located around -460, -425, and -405 with respect to the ATG. RNase protection analysis indicated that once the negative regulatory sequences were deleted, transcription in NIH 3T3 cells initiated from the same transcription initiation sites normally utilized in spermatocytes, approximately 280 nucleotides upstream of the ATG. Deletions beyond the spermatocyte promoter, however, allowed transcription initiation from progressively downstream c-mos sequences. Deletion or mutation of sequences surrounding the oocyte promoter at -53 also had little effect on expression of c-mos constructs in NIH 3T3 cells. Therefore, the major determinant of c-mos expression in NIH 3T3 cells was removal of the negative regulatory sequence rather than the utilization of a unique promoter. The c-mos negative regulatory sequences thus appear to play a significant role in tissue-specific c-mos expression by inhibiting transcription in somatic cells.

Original languageEnglish
Pages (from-to)2029-2036
Number of pages8
JournalMolecular and Cellular Biology
Volume12
Issue number5
Publication statusPublished - May 1992

Fingerprint

NIH 3T3 Cells
Spermatocytes
Nucleotides
BALB 3T3 Cells
Sequence Deletion
5' Flanking Region
Transcription Initiation Site
Pheochromocytoma
Ribonucleases
Site-Directed Mutagenesis
Oocytes
Carcinoma
Lung

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Identification of a negative regulatory element that inhibits c-mos transcription in somatic cells. / Zinkel, Sandra S.; Pal, Subrata K.; Szeberényi, J.; Cooper, Geoffrey M.

In: Molecular and Cellular Biology, Vol. 12, No. 5, 05.1992, p. 2029-2036.

Research output: Contribution to journalArticle

Zinkel, Sandra S. ; Pal, Subrata K. ; Szeberényi, J. ; Cooper, Geoffrey M. / Identification of a negative regulatory element that inhibits c-mos transcription in somatic cells. In: Molecular and Cellular Biology. 1992 ; Vol. 12, No. 5. pp. 2029-2036.
@article{746dddecfe974cb0aaac02e3df96cf62,
title = "Identification of a negative regulatory element that inhibits c-mos transcription in somatic cells",
abstract = "We have used transient expression assays to identify a cis-acting region in the 5′ flanking sequence of murine c-mos which, when deleted, allows expression from the c-mos promoter in NIH 3T3 cells. This negative regulatory sequence, located 400 to 500 nucleotides upstream of the c-mos ATG, also inhibited expression from a heterologous promoter. In addition to NIH 3T3 cells, the c-mos negative regulatory sequence was active in BALB/3T3 cells, PC12 rat pheochromocytoma cells, and A549 human lung carcinoma cells. Site-specific mutagenesis identified three possibly interacting regions that were involved in negative regulatory activity, located around -460, -425, and -405 with respect to the ATG. RNase protection analysis indicated that once the negative regulatory sequences were deleted, transcription in NIH 3T3 cells initiated from the same transcription initiation sites normally utilized in spermatocytes, approximately 280 nucleotides upstream of the ATG. Deletions beyond the spermatocyte promoter, however, allowed transcription initiation from progressively downstream c-mos sequences. Deletion or mutation of sequences surrounding the oocyte promoter at -53 also had little effect on expression of c-mos constructs in NIH 3T3 cells. Therefore, the major determinant of c-mos expression in NIH 3T3 cells was removal of the negative regulatory sequence rather than the utilization of a unique promoter. The c-mos negative regulatory sequences thus appear to play a significant role in tissue-specific c-mos expression by inhibiting transcription in somatic cells.",
author = "Zinkel, {Sandra S.} and Pal, {Subrata K.} and J. Szeber{\'e}nyi and Cooper, {Geoffrey M.}",
year = "1992",
month = "5",
language = "English",
volume = "12",
pages = "2029--2036",
journal = "Molecular and Cellular Biology",
issn = "0270-7306",
publisher = "American Society for Microbiology",
number = "5",

}

TY - JOUR

T1 - Identification of a negative regulatory element that inhibits c-mos transcription in somatic cells

AU - Zinkel, Sandra S.

AU - Pal, Subrata K.

AU - Szeberényi, J.

AU - Cooper, Geoffrey M.

PY - 1992/5

Y1 - 1992/5

N2 - We have used transient expression assays to identify a cis-acting region in the 5′ flanking sequence of murine c-mos which, when deleted, allows expression from the c-mos promoter in NIH 3T3 cells. This negative regulatory sequence, located 400 to 500 nucleotides upstream of the c-mos ATG, also inhibited expression from a heterologous promoter. In addition to NIH 3T3 cells, the c-mos negative regulatory sequence was active in BALB/3T3 cells, PC12 rat pheochromocytoma cells, and A549 human lung carcinoma cells. Site-specific mutagenesis identified three possibly interacting regions that were involved in negative regulatory activity, located around -460, -425, and -405 with respect to the ATG. RNase protection analysis indicated that once the negative regulatory sequences were deleted, transcription in NIH 3T3 cells initiated from the same transcription initiation sites normally utilized in spermatocytes, approximately 280 nucleotides upstream of the ATG. Deletions beyond the spermatocyte promoter, however, allowed transcription initiation from progressively downstream c-mos sequences. Deletion or mutation of sequences surrounding the oocyte promoter at -53 also had little effect on expression of c-mos constructs in NIH 3T3 cells. Therefore, the major determinant of c-mos expression in NIH 3T3 cells was removal of the negative regulatory sequence rather than the utilization of a unique promoter. The c-mos negative regulatory sequences thus appear to play a significant role in tissue-specific c-mos expression by inhibiting transcription in somatic cells.

AB - We have used transient expression assays to identify a cis-acting region in the 5′ flanking sequence of murine c-mos which, when deleted, allows expression from the c-mos promoter in NIH 3T3 cells. This negative regulatory sequence, located 400 to 500 nucleotides upstream of the c-mos ATG, also inhibited expression from a heterologous promoter. In addition to NIH 3T3 cells, the c-mos negative regulatory sequence was active in BALB/3T3 cells, PC12 rat pheochromocytoma cells, and A549 human lung carcinoma cells. Site-specific mutagenesis identified three possibly interacting regions that were involved in negative regulatory activity, located around -460, -425, and -405 with respect to the ATG. RNase protection analysis indicated that once the negative regulatory sequences were deleted, transcription in NIH 3T3 cells initiated from the same transcription initiation sites normally utilized in spermatocytes, approximately 280 nucleotides upstream of the ATG. Deletions beyond the spermatocyte promoter, however, allowed transcription initiation from progressively downstream c-mos sequences. Deletion or mutation of sequences surrounding the oocyte promoter at -53 also had little effect on expression of c-mos constructs in NIH 3T3 cells. Therefore, the major determinant of c-mos expression in NIH 3T3 cells was removal of the negative regulatory sequence rather than the utilization of a unique promoter. The c-mos negative regulatory sequences thus appear to play a significant role in tissue-specific c-mos expression by inhibiting transcription in somatic cells.

UR - http://www.scopus.com/inward/record.url?scp=0026666972&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026666972&partnerID=8YFLogxK

M3 - Article

C2 - 1533271

AN - SCOPUS:0026666972

VL - 12

SP - 2029

EP - 2036

JO - Molecular and Cellular Biology

JF - Molecular and Cellular Biology

SN - 0270-7306

IS - 5

ER -