Recombinant interferon-γ differently affects DQA and DRA gene expression in two human melanoma clones

Transcriptional and posttranscriptional regulation

C. Castelli, M. Sensi, G. Parmiani

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

In a previous study, two human melanoma HLA class-II-negative clones have been shown to respond differently to recombinant interferon gamma (rINF-γ). In fact, in clone 9229/18, rINF-γ treatment led to a coordinate expression of both DR and DQ genes, whereas in clone 9229/5, a high increase in steady-state mRNA levels and cell surface antigen expression could be observed for DR but not for DQ genes. The molecular mechanism underlying such a different behaviour was investigated and DQA gene regulaton was studied both at the transcriptional and posttranscriptional level. Nuclear run-on experiments were performed on 9229/5 and 9229/18 clones. Treatment with rINF-γ at 1,000 U/ml for 24 h led to a coordinate transcriptional activation of DRA and DQA genes in 9229/18 clone, whereas in clone 9229/5 it strongly augmented the rate of transcription of DRA but not DQA genes. For all class II genes studied, both melanoma clones showed a basal rate of transcription that never led to a mature cyptoplasmatic mRNA. To study wether posttranscriptional mechanisms could affect DQA mRNA stability, a comparison was performed between mRNA turnover in 9229/18 cells treated with actinomycin D and actinomycin D plus cyclohexamide following rINF-γ treatment. In the absence of protein synthesis, the t( 1/2 ) of specific DQA mRNA was largely reduced, showing that a short-lived protein is required to stabilize human DQA mRNA in melanoma cells. Our results indicate that DQA gene is subjected to a tight regulation, acting both at the transcriptional and posttranscriptional levels and that DQA and DRA genes can be differentially regulated at the transcriptional level by rINF-γ in a melanoma clone such as 9229/5.

Original languageEnglish
Pages (from-to)38-48
Number of pages11
JournalExperimental and Clinical Immunogenetics
Volume8
Issue number1
Publication statusPublished - 1991

Fingerprint

Interferons
Melanoma
Clone Cells
Gene Expression
Interferon-gamma
Genes
Messenger RNA
Dactinomycin
MHC Class II Genes
RNA Stability
Surface Antigens
Transcriptional Activation
Proteins

ASJC Scopus subject areas

  • Immunology
  • Genetics(clinical)
  • Genetics

Cite this

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title = "Recombinant interferon-γ differently affects DQA and DRA gene expression in two human melanoma clones: Transcriptional and posttranscriptional regulation",
abstract = "In a previous study, two human melanoma HLA class-II-negative clones have been shown to respond differently to recombinant interferon gamma (rINF-γ). In fact, in clone 9229/18, rINF-γ treatment led to a coordinate expression of both DR and DQ genes, whereas in clone 9229/5, a high increase in steady-state mRNA levels and cell surface antigen expression could be observed for DR but not for DQ genes. The molecular mechanism underlying such a different behaviour was investigated and DQA gene regulaton was studied both at the transcriptional and posttranscriptional level. Nuclear run-on experiments were performed on 9229/5 and 9229/18 clones. Treatment with rINF-γ at 1,000 U/ml for 24 h led to a coordinate transcriptional activation of DRA and DQA genes in 9229/18 clone, whereas in clone 9229/5 it strongly augmented the rate of transcription of DRA but not DQA genes. For all class II genes studied, both melanoma clones showed a basal rate of transcription that never led to a mature cyptoplasmatic mRNA. To study wether posttranscriptional mechanisms could affect DQA mRNA stability, a comparison was performed between mRNA turnover in 9229/18 cells treated with actinomycin D and actinomycin D plus cyclohexamide following rINF-γ treatment. In the absence of protein synthesis, the t( 1/2 ) of specific DQA mRNA was largely reduced, showing that a short-lived protein is required to stabilize human DQA mRNA in melanoma cells. Our results indicate that DQA gene is subjected to a tight regulation, acting both at the transcriptional and posttranscriptional levels and that DQA and DRA genes can be differentially regulated at the transcriptional level by rINF-γ in a melanoma clone such as 9229/5.",
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T1 - Recombinant interferon-γ differently affects DQA and DRA gene expression in two human melanoma clones

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AU - Castelli, C.

AU - Sensi, M.

AU - Parmiani, G.

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N2 - In a previous study, two human melanoma HLA class-II-negative clones have been shown to respond differently to recombinant interferon gamma (rINF-γ). In fact, in clone 9229/18, rINF-γ treatment led to a coordinate expression of both DR and DQ genes, whereas in clone 9229/5, a high increase in steady-state mRNA levels and cell surface antigen expression could be observed for DR but not for DQ genes. The molecular mechanism underlying such a different behaviour was investigated and DQA gene regulaton was studied both at the transcriptional and posttranscriptional level. Nuclear run-on experiments were performed on 9229/5 and 9229/18 clones. Treatment with rINF-γ at 1,000 U/ml for 24 h led to a coordinate transcriptional activation of DRA and DQA genes in 9229/18 clone, whereas in clone 9229/5 it strongly augmented the rate of transcription of DRA but not DQA genes. For all class II genes studied, both melanoma clones showed a basal rate of transcription that never led to a mature cyptoplasmatic mRNA. To study wether posttranscriptional mechanisms could affect DQA mRNA stability, a comparison was performed between mRNA turnover in 9229/18 cells treated with actinomycin D and actinomycin D plus cyclohexamide following rINF-γ treatment. In the absence of protein synthesis, the t( 1/2 ) of specific DQA mRNA was largely reduced, showing that a short-lived protein is required to stabilize human DQA mRNA in melanoma cells. Our results indicate that DQA gene is subjected to a tight regulation, acting both at the transcriptional and posttranscriptional levels and that DQA and DRA genes can be differentially regulated at the transcriptional level by rINF-γ in a melanoma clone such as 9229/5.

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