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Regulation in-vitro of an l-cam enhancer by homeobox genes hoxd9 and hnf-1

Academic Article
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Overview

authors

  • Goomer, R. S.
  • Holst, B. D.
  • Wood, I. C.
  • Jones, F. S.
  • Edelman, Gerald

publication date

  • August 1994

journal

  • Proceedings of the National Academy of Sciences of the United States of America  Journal

abstract

  • Previous studies have shown that in vitro expression of the neural cell adhesion molecule (N-CAM) can be regulated by the products of homeobox genes HoxB9, -B8, and -C6. N-CAM is a Ca(2+)-independent immunoglobulin-related CAM that plays an important role in neural development. In the present study, we investigated whether the liver cell adhesion molecule (L-CAM) a member of the Ca(2+)-dependent CAM family (cadherins) is also regulated by homeobox-containing genes. In transient cotransfection experiments of NIH 3T3 cells, we observed that both HoxD9 and liver-enriched POU-homeodomain transcription factor, HNF-1, activated chloramphenicol acetyltransferase gene reporter constructs containing the L-CAM promoter and an enhancer present in the second intron of the chicken L-CAM gene. Using electrophoretic mobility-shift assays, we found that components of cell extracts from NIH 3T3 cells transfected with HoxD9 bound to a small region of the L-CAM enhancer having a consensus sequence that is a putative binding site for HNF-1. Components of extracts from the chicken hepatoma cell line LMH that had been transfected with an HNF-1 expression vector also bound to this same site. In nuclear run-on experiments with nuclei from LMH cells that were transfected with expression vectors for HoxD9 or HNF-1, L-CAM RNA levels were increased 33-fold and 4-fold respectively. Using the same run-on procedure, it was confirmed that nuclei prepared from normal embryonic chicken liver cells expressed the RNAs for HoxD9, HNF-1, and L-CAM. Taken together with previous observations, these data raise the possibility that homeobox-containing genes will have a widespread role in the place-dependent expression of CAMs belonging both to immunoglobulin-related and to cadherin families.

subject areas

  • 3T3 Cells
  • Animals
  • Base Sequence
  • Binding Sites
  • Cadherins
  • Chickens
  • Consensus Sequence
  • DNA-Binding Proteins
  • Enhancer Elements, Genetic
  • Gene Expression Regulation
  • Genes, Homeobox
  • Hepatocyte Nuclear Factor 1
  • Hepatocyte Nuclear Factor 1-alpha
  • Hepatocyte Nuclear Factor 1-beta
  • Introns
  • Liver
  • Mice
  • Molecular Sequence Data
  • Neoplasm Proteins
  • Nuclear Proteins
  • Oligonucleotide Probes
  • Promoter Regions, Genetic
  • Transcription Factors
  • Transfection
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Research

keywords

  • CADHERINS
  • GENE EXPRESSION
  • HOMEODOMAIN PROTEINS
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Identity

International Standard Serial Number (ISSN)

  • 0027-8424

Digital Object Identifier (DOI)

  • 10.1073/pnas.91.17.7985

PubMed ID

  • 7914699
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Additional Document Info

start page

  • 7985

end page

  • 7989

volume

  • 91

issue

  • 17

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