Cell type-specific activity of the N-myc promoter in human neuroblastoma cells is mediated by a downstream silencer

K. A. Woodruff, Joseph D Rosenblatt, T. B. Moore, R. H. Medzoyan, D. S M Pai, J. L. Noland, J. M. Yamashiro, R. K. Wada

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The N-myc oncogene is actively transcribed in many neuroblastoma tumors, but is not expressed in mature, normal tissue of any type. Chloramphenicol acetyl transferase (CAT) assays of constructs containing N-myc sequence transfected into N-myc expressing LA-N-5 neuroblastoma cells or non-expressing HeLa carcinoma cells have revealed a 201 base pair (bp) regulatory region mediating the cell type-specific activity of the promoter. While located downstream from 5' mRNA cap sites, the region appears to function by preventing transcriptional initiation. This downstream region is capable of suppressing promoter activity independently of position, and contains an element having 100% homology with the 9 bp consensus sequence of a transcriptional silencer found in the upstream region of the lysozyme gene. DNA gel retardation assays have shown that this sequence is involved in a specific DNA-protein interaction with nuclear extract from HeLa cells that is distinct from that occurring with extract from LA-N-5 cells. These results suggest that the N-myc promoter's cell type-specific activity is regulated by a downstream silencer, and that differential binding of regulatory protein from that present in non-expressing cells may result in the constitutive N-myc expression seen in neuroblastoma.

Original languageEnglish
Pages (from-to)1335-1341
Number of pages7
JournalOncogene
Volume10
Issue number7
StatePublished - Jan 1 1995
Externally publishedYes

Fingerprint

Neuroblastoma
HeLa Cells
Base Pairing
myc Genes
Nucleic Acid Regulatory Sequences
DNA
Consensus Sequence
Electrophoretic Mobility Shift Assay
Chloramphenicol
Muramidase
Transferases
Carrier Proteins
Carcinoma
Messenger RNA
Genes
Neoplasms
Proteins

Keywords

  • Downstream silencer
  • N-myc
  • Neuroblastoma

ASJC Scopus subject areas

  • Cancer Research
  • Genetics
  • Molecular Biology

Cite this

Woodruff, K. A., Rosenblatt, J. D., Moore, T. B., Medzoyan, R. H., Pai, D. S. M., Noland, J. L., ... Wada, R. K. (1995). Cell type-specific activity of the N-myc promoter in human neuroblastoma cells is mediated by a downstream silencer. Oncogene, 10(7), 1335-1341.

Cell type-specific activity of the N-myc promoter in human neuroblastoma cells is mediated by a downstream silencer. / Woodruff, K. A.; Rosenblatt, Joseph D; Moore, T. B.; Medzoyan, R. H.; Pai, D. S M; Noland, J. L.; Yamashiro, J. M.; Wada, R. K.

In: Oncogene, Vol. 10, No. 7, 01.01.1995, p. 1335-1341.

Research output: Contribution to journalArticle

Woodruff, KA, Rosenblatt, JD, Moore, TB, Medzoyan, RH, Pai, DSM, Noland, JL, Yamashiro, JM & Wada, RK 1995, 'Cell type-specific activity of the N-myc promoter in human neuroblastoma cells is mediated by a downstream silencer', Oncogene, vol. 10, no. 7, pp. 1335-1341.
Woodruff KA, Rosenblatt JD, Moore TB, Medzoyan RH, Pai DSM, Noland JL et al. Cell type-specific activity of the N-myc promoter in human neuroblastoma cells is mediated by a downstream silencer. Oncogene. 1995 Jan 1;10(7):1335-1341.
Woodruff, K. A. ; Rosenblatt, Joseph D ; Moore, T. B. ; Medzoyan, R. H. ; Pai, D. S M ; Noland, J. L. ; Yamashiro, J. M. ; Wada, R. K. / Cell type-specific activity of the N-myc promoter in human neuroblastoma cells is mediated by a downstream silencer. In: Oncogene. 1995 ; Vol. 10, No. 7. pp. 1335-1341.
@article{4266b92f711349c8b0490b0cc82b1c9f,
title = "Cell type-specific activity of the N-myc promoter in human neuroblastoma cells is mediated by a downstream silencer",
abstract = "The N-myc oncogene is actively transcribed in many neuroblastoma tumors, but is not expressed in mature, normal tissue of any type. Chloramphenicol acetyl transferase (CAT) assays of constructs containing N-myc sequence transfected into N-myc expressing LA-N-5 neuroblastoma cells or non-expressing HeLa carcinoma cells have revealed a 201 base pair (bp) regulatory region mediating the cell type-specific activity of the promoter. While located downstream from 5' mRNA cap sites, the region appears to function by preventing transcriptional initiation. This downstream region is capable of suppressing promoter activity independently of position, and contains an element having 100{\%} homology with the 9 bp consensus sequence of a transcriptional silencer found in the upstream region of the lysozyme gene. DNA gel retardation assays have shown that this sequence is involved in a specific DNA-protein interaction with nuclear extract from HeLa cells that is distinct from that occurring with extract from LA-N-5 cells. These results suggest that the N-myc promoter's cell type-specific activity is regulated by a downstream silencer, and that differential binding of regulatory protein from that present in non-expressing cells may result in the constitutive N-myc expression seen in neuroblastoma.",
keywords = "Downstream silencer, N-myc, Neuroblastoma",
author = "Woodruff, {K. A.} and Rosenblatt, {Joseph D} and Moore, {T. B.} and Medzoyan, {R. H.} and Pai, {D. S M} and Noland, {J. L.} and Yamashiro, {J. M.} and Wada, {R. K.}",
year = "1995",
month = "1",
day = "1",
language = "English",
volume = "10",
pages = "1335--1341",
journal = "Oncogene",
issn = "0950-9232",
publisher = "Nature Publishing Group",
number = "7",

}

TY - JOUR

T1 - Cell type-specific activity of the N-myc promoter in human neuroblastoma cells is mediated by a downstream silencer

AU - Woodruff, K. A.

AU - Rosenblatt, Joseph D

AU - Moore, T. B.

AU - Medzoyan, R. H.

AU - Pai, D. S M

AU - Noland, J. L.

AU - Yamashiro, J. M.

AU - Wada, R. K.

PY - 1995/1/1

Y1 - 1995/1/1

N2 - The N-myc oncogene is actively transcribed in many neuroblastoma tumors, but is not expressed in mature, normal tissue of any type. Chloramphenicol acetyl transferase (CAT) assays of constructs containing N-myc sequence transfected into N-myc expressing LA-N-5 neuroblastoma cells or non-expressing HeLa carcinoma cells have revealed a 201 base pair (bp) regulatory region mediating the cell type-specific activity of the promoter. While located downstream from 5' mRNA cap sites, the region appears to function by preventing transcriptional initiation. This downstream region is capable of suppressing promoter activity independently of position, and contains an element having 100% homology with the 9 bp consensus sequence of a transcriptional silencer found in the upstream region of the lysozyme gene. DNA gel retardation assays have shown that this sequence is involved in a specific DNA-protein interaction with nuclear extract from HeLa cells that is distinct from that occurring with extract from LA-N-5 cells. These results suggest that the N-myc promoter's cell type-specific activity is regulated by a downstream silencer, and that differential binding of regulatory protein from that present in non-expressing cells may result in the constitutive N-myc expression seen in neuroblastoma.

AB - The N-myc oncogene is actively transcribed in many neuroblastoma tumors, but is not expressed in mature, normal tissue of any type. Chloramphenicol acetyl transferase (CAT) assays of constructs containing N-myc sequence transfected into N-myc expressing LA-N-5 neuroblastoma cells or non-expressing HeLa carcinoma cells have revealed a 201 base pair (bp) regulatory region mediating the cell type-specific activity of the promoter. While located downstream from 5' mRNA cap sites, the region appears to function by preventing transcriptional initiation. This downstream region is capable of suppressing promoter activity independently of position, and contains an element having 100% homology with the 9 bp consensus sequence of a transcriptional silencer found in the upstream region of the lysozyme gene. DNA gel retardation assays have shown that this sequence is involved in a specific DNA-protein interaction with nuclear extract from HeLa cells that is distinct from that occurring with extract from LA-N-5 cells. These results suggest that the N-myc promoter's cell type-specific activity is regulated by a downstream silencer, and that differential binding of regulatory protein from that present in non-expressing cells may result in the constitutive N-myc expression seen in neuroblastoma.

KW - Downstream silencer

KW - N-myc

KW - Neuroblastoma

UR - http://www.scopus.com/inward/record.url?scp=0028924346&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028924346&partnerID=8YFLogxK

M3 - Article

VL - 10

SP - 1335

EP - 1341

JO - Oncogene

JF - Oncogene

SN - 0950-9232

IS - 7

ER -