Expression of the cystic fibrosis transmembrane conductance regulator gene in cells of non-epithelial origin

Kunihiko Yoshimura, Hidenori Nakamura, Bruce C. Trapnell, Chin shyan Chu, Wilfeuwnd Dakemans, Andrea Pavirani, Jean pierre Lecocq, Ronald Crystal

Research output: Contribution to journalArticle

104 Citations (Scopus)

Abstract

Consistent with the fact that the clinical disorder cystic fibrosis (CF) is manifested on epithelial surfaces, active transcription of the CF transmembrane conductance regulator (CFTR) gene and CFTR mRNA transcripts are detectable in a variety of epithelial cells, suggesting CFTR gene expression might be epithelial cell-specific. However, analysis of the CFTR gene promoter suggests it is a housekeeping gene, implying more widespread expression than only in epithelial cells. To evaluate the latter hypothesis, various human cells of non-epithelial origin, including lung fibroblasts, U-937 histiocytic lymphoma cells, K-562 erythroleukemia cells, HL-60 promyelocytic leukemia cells as well as freshly isolated blood lymphocytes, neutrophils, monocytes, and alveolar macrophages were examined for CFTR gene expression. Although Northern analysis failed to show CFTR mRNA transcripts in these cells, amplification of mRNA (after conversion to cDNA) by polymerase chain reaction combined with Southern analysis demonstrated the presence of CFTR mRNA transcripts at low levels in all cells evaluated except HL-60 cells. Comparative quantitative analysis showed fibroblasts contained 200 - 400 fold less CFTR mRNA transcripts than the T84 and HT-29 colon carcinoma epithelial cell lines, but had similar levels of CFTR transcripts to those of other epithelial cell lines. Nuclear transcription runon analyses demonstrated very low level CFTR gene transcription in fibroblasts and U-937 cells, similar to that of other epithelial cells, but lower than the T84 and HT-29 colon carcinoma cell lines. Interestingly, while chromatin DNA of fibroblasts had no DNase I hypersensitivity sites in the 5′ flanking region of the CFTR gene, HT-29 chromatin DNA exhibited four DNase I accessible sites in the same region, suggesting that these sites may be related to more active transcription of the CFTR gene in the intestinal epithelial cells than in fibroblasts.

Original languageEnglish
Pages (from-to)5417-5423
Number of pages7
JournalNucleic Acids Research
Volume19
Issue number19
DOIs
Publication statusPublished - 11 Oct 1991
Externally publishedYes

Fingerprint

Cystic Fibrosis Transmembrane Conductance Regulator
Regulator Genes
Epithelial Cells
Fibroblasts
Messenger RNA
HL-60 Cells
Deoxyribonuclease I
Cell Line
Chromatin
Colon
Carcinoma
Gene Expression
Leukemia, Erythroblastic, Acute
Lymphoma, Large B-Cell, Diffuse
5' Flanking Region
DNA
Essential Genes
Alveolar Macrophages
Cystic Fibrosis
Monocytes

ASJC Scopus subject areas

  • Genetics

Cite this

Expression of the cystic fibrosis transmembrane conductance regulator gene in cells of non-epithelial origin. / Yoshimura, Kunihiko; Nakamura, Hidenori; Trapnell, Bruce C.; Chu, Chin shyan; Dakemans, Wilfeuwnd; Pavirani, Andrea; Lecocq, Jean pierre; Crystal, Ronald.

In: Nucleic Acids Research, Vol. 19, No. 19, 11.10.1991, p. 5417-5423.

Research output: Contribution to journalArticle

Yoshimura, K, Nakamura, H, Trapnell, BC, Chu, CS, Dakemans, W, Pavirani, A, Lecocq, JP & Crystal, R 1991, 'Expression of the cystic fibrosis transmembrane conductance regulator gene in cells of non-epithelial origin', Nucleic Acids Research, vol. 19, no. 19, pp. 5417-5423. https://doi.org/10.1093/nar/19.19.5417
Yoshimura, Kunihiko ; Nakamura, Hidenori ; Trapnell, Bruce C. ; Chu, Chin shyan ; Dakemans, Wilfeuwnd ; Pavirani, Andrea ; Lecocq, Jean pierre ; Crystal, Ronald. / Expression of the cystic fibrosis transmembrane conductance regulator gene in cells of non-epithelial origin. In: Nucleic Acids Research. 1991 ; Vol. 19, No. 19. pp. 5417-5423.
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