Transcription analysis, physical mapping, and molecular characterization of a nonclassical human leukocyte antigen class I gene

Michael J. Chorney, Ikuhisa Sawada, Gerald A. Gillespie, Rakesh Srivastava, Julian Pan, Sherman M. Weissman

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

The human major histocompatibility complex contains approximately 20 class I genes, pseudogenes, and gene fragments. These include the genes for the three major transplantation antigens, HLA-A, HLA-B, and HLA-C, as well as a number of other genes or pseudogenes of unknown biological significance. Most of the latter have C+G-rich sequences in their 5′ ends that are unmethylated in the B-lymphoblastoid cell line 3.1.0. We investigated one of these genes, HLA-H, in more detail. The gene is, overall, strongly homologous in sequence to HLA-A but differs in several potentially significant ways, including changes in conserved promoter sequences, a single-base deletion producing a translation termination codon in exon 4, and a region of sequence divergence downstream of the transcribed portion of the gene. Nevertheless, mouse L cells transfected with the gene accumulated small amounts of apparently full-length polyadenylated RNA. A portion of this RNA begins at the transcription site predicted by analogy to certain class I cDNA clones, while another portion appears to begin shortly upstream. L cells transfected with a hybrid gene containing the first three exons of HLA-H and the last five exons of HLA-B27 accumulated full-length HLA transcripts at the same level as cells transfected with an HLA-B27 gene; both levels are at least 15- to 20-fold higher than that directed by HLA-H alone. In addition, we isolated a cDNA clone for HLA-H that contains a portion of intron 3 attached to a normally spliced sequence comprising exons 4 through 8. These results suggest that low levels of translatable mRNA for the truncated class I heavy chain encoded by HLA-H are produced under physiologic circumstances and that sequences 3′ of intron 3 decrease the levels of stable transcripts.

Original languageEnglish (US)
Pages (from-to)243-253
Number of pages11
JournalMolecular and cellular biology
Volume10
Issue number1
DOIs
StatePublished - Jan 1 1990

Fingerprint

MHC Class I Genes
HLA Antigens
Genes
Exons
HLA-B27 Antigen
Pseudogenes
HLA-A Antigens
Introns
Complementary DNA
Clone Cells
HLA-C Antigens
Messenger RNA
Histocompatibility Antigens
HLA-B Antigens
Terminator Codon
Conserved Sequence
Sequence Homology
Major Histocompatibility Complex
RNA
Cell Line

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Chorney, Michael J. ; Sawada, Ikuhisa ; Gillespie, Gerald A. ; Srivastava, Rakesh ; Pan, Julian ; Weissman, Sherman M. / Transcription analysis, physical mapping, and molecular characterization of a nonclassical human leukocyte antigen class I gene. In: Molecular and cellular biology. 1990 ; Vol. 10, No. 1. pp. 243-253.
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abstract = "The human major histocompatibility complex contains approximately 20 class I genes, pseudogenes, and gene fragments. These include the genes for the three major transplantation antigens, HLA-A, HLA-B, and HLA-C, as well as a number of other genes or pseudogenes of unknown biological significance. Most of the latter have C+G-rich sequences in their 5′ ends that are unmethylated in the B-lymphoblastoid cell line 3.1.0. We investigated one of these genes, HLA-H, in more detail. The gene is, overall, strongly homologous in sequence to HLA-A but differs in several potentially significant ways, including changes in conserved promoter sequences, a single-base deletion producing a translation termination codon in exon 4, and a region of sequence divergence downstream of the transcribed portion of the gene. Nevertheless, mouse L cells transfected with the gene accumulated small amounts of apparently full-length polyadenylated RNA. A portion of this RNA begins at the transcription site predicted by analogy to certain class I cDNA clones, while another portion appears to begin shortly upstream. L cells transfected with a hybrid gene containing the first three exons of HLA-H and the last five exons of HLA-B27 accumulated full-length HLA transcripts at the same level as cells transfected with an HLA-B27 gene; both levels are at least 15- to 20-fold higher than that directed by HLA-H alone. In addition, we isolated a cDNA clone for HLA-H that contains a portion of intron 3 attached to a normally spliced sequence comprising exons 4 through 8. These results suggest that low levels of translatable mRNA for the truncated class I heavy chain encoded by HLA-H are produced under physiologic circumstances and that sequences 3′ of intron 3 decrease the levels of stable transcripts.",
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Transcription analysis, physical mapping, and molecular characterization of a nonclassical human leukocyte antigen class I gene. / Chorney, Michael J.; Sawada, Ikuhisa; Gillespie, Gerald A.; Srivastava, Rakesh; Pan, Julian; Weissman, Sherman M.

In: Molecular and cellular biology, Vol. 10, No. 1, 01.01.1990, p. 243-253.

Research output: Contribution to journalArticle

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