The 3' untranslated region plays a role in the targeting of metallothionein-I mRNA to the perinuclear cytoplasm and cytoskeletal-bound polysomes

P Mahon, K Partridge, J H Beattie, L A Glover, J E Hesketh

Research output: Contribution to journalArticle

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Abstract

The mechanism of localisation of metallothionein-I (MT-I) mRNA was studied in transfected cells by in situ hybridisation and cell fractionation. Hepatoma cells were transfected with the 5'-untranslated region and coding region of the beta-globin gene alone or linked to either the beta-globin 3'-untranslated region (3'-UTR) or the MT-I 3'-UTR. The wild-type beta-globin mRNA and the beta-globin mRNA lacking its native 3'-UTR were present in free and cytoskeletal-bound polysomes to a similar extent and showed no localisation. Chimaeric globin-metallothionein transcripts were significantly enriched in cytoskeletal-bound polysomes and were localised in the perinuclear cytoplasm. Chimaeric globin-metallothionein and wild-type globin transcripts were of similar stability. Chinese Hamster Ovary cells were transfected with constructs in which the MT-I 5'-untranslated region and coding sequences were linked to either the endogenous 3'-UTR or the glutathione peroxidase 3'-UTR. Wild-type MT-I transcripts were localised in the perinuclear cytoplasm but the chimaeric MT-I-glutathione peroxidase transcripts showed no distinct localisation. The results indicate that the 3'-UTR of MT-I mRNA contains a localisation signal which promotes both the association of the mRNA with the cytoskeleton and its perinuclear localisation. (C) 1997 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)153-162
Number of pages10
JournalBiochimica et Biophysica Acta. Molecular Cell Research
Volume1358
Issue number2
Publication statusPublished - 11 Sep 1997

Keywords

  • 3'-untranslated region
  • metallothionein
  • mRNA localisation
  • cytoskeleton
  • polyribosome
  • translation
  • MESSENGER-RNA LOCALIZATION
  • C-MYC
  • XENOPUS OOCYTES
  • RAT HEPATOCYTES
  • GENE-EXPRESSION
  • HELA-CELLS
  • SEQUENCES
  • PROTEINS
  • FIBROBLASTS

Cite this

The 3' untranslated region plays a role in the targeting of metallothionein-I mRNA to the perinuclear cytoplasm and cytoskeletal-bound polysomes. / Mahon, P ; Partridge, K ; Beattie, J H ; Glover, L A ; Hesketh, J E .

In: Biochimica et Biophysica Acta. Molecular Cell Research, Vol. 1358, No. 2, 11.09.1997, p. 153-162.

Research output: Contribution to journalArticle

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AB - The mechanism of localisation of metallothionein-I (MT-I) mRNA was studied in transfected cells by in situ hybridisation and cell fractionation. Hepatoma cells were transfected with the 5'-untranslated region and coding region of the beta-globin gene alone or linked to either the beta-globin 3'-untranslated region (3'-UTR) or the MT-I 3'-UTR. The wild-type beta-globin mRNA and the beta-globin mRNA lacking its native 3'-UTR were present in free and cytoskeletal-bound polysomes to a similar extent and showed no localisation. Chimaeric globin-metallothionein transcripts were significantly enriched in cytoskeletal-bound polysomes and were localised in the perinuclear cytoplasm. Chimaeric globin-metallothionein and wild-type globin transcripts were of similar stability. Chinese Hamster Ovary cells were transfected with constructs in which the MT-I 5'-untranslated region and coding sequences were linked to either the endogenous 3'-UTR or the glutathione peroxidase 3'-UTR. Wild-type MT-I transcripts were localised in the perinuclear cytoplasm but the chimaeric MT-I-glutathione peroxidase transcripts showed no distinct localisation. The results indicate that the 3'-UTR of MT-I mRNA contains a localisation signal which promotes both the association of the mRNA with the cytoskeleton and its perinuclear localisation. (C) 1997 Elsevier Science B.V.

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KW - mRNA localisation

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JO - Biochimica et Biophysica Acta. Molecular Cell Research

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