Analysis of full length ADAMTS6 transcript reveals alternative splicing and a role for the 5' untranslated region in translational control
| Autor: | Jenny L. Lindrop, Debra J. Bevitt, Norman McKie, Zheng Li, Michael P. Clarke, Michael D. Barker |
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| Rok vydání: | 2004 |
| Předmět: |
Untranslated region
DNA Complementary Five prime untranslated region Transcription Genetic Recombinant Fusion Proteins Molecular Sequence Data Codon Initiator Biology Cell Line Open Reading Frames ADAMTS Proteins Start codon Catalytic Domain Sequence Homology Nucleic Acid Genetics Humans Point Mutation Amino Acid Sequence RNA Messenger Luciferases Messenger RNA Reporter gene Binding Sites Base Sequence Sequence Homology Amino Acid Reverse Transcriptase Polymerase Chain Reaction Tumor Necrosis Factor-alpha Alternative splicing General Medicine Sequence Analysis DNA Molecular biology Isoenzymes Open reading frame Internal ribosome entry site ADAM Proteins Alternative Splicing Gene Expression Regulation Protein Biosynthesis 5' Untranslated Regions Ribosomes Sequence Alignment |
| Zdroj: | Gene. 359 |
| ISSN: | 0378-1119 |
| Popis: | The ADAMTS (A Disintegrin and metalloproteinase with thrombospondin-1 type repeats) family of enzymes have been implicated in turnover of extracellular matrix. We previously showed that levels of ADAMTS6 mRNA in ARPE-19 cells were markedly increased following treatment with tumour necrosis factor alpha (TNFα). This study shows that the ADAMTS6 transcript contains unusually large untranslated regions (UTRs) at both the 5′ and 3′end. The 5′UTR contains 11 AUG codons upstream of the predicted ADAMTS6 start codon and potently inhibits translation of a downstream reporter gene. However some translation can be restored by truncating the 5′UTR from the 5′end. The 5′UTR was tested for internal ribosome entry site activity using a bicistronic luciferase reporter plasmid, but none was detected. Using the 5′ and 3′UTR sequences to screen the GenBank database we identified a full length ADAMTS6 cDNA of 7262 bp. This transcript is alternatively spliced at the 3′end of the open reading frame (ORF), resulting in an extended ORF containing 3 additional tsp-1 type repeats. Quantitative RT-PCR showed that the long and short form of the ADAMTS6 ORF are co-expressed in ARPE-19 cells, but the relative levels of the two forms is modulated by TNFα. The region of the transcript encoding the catalytic domain also contains several notable differences compared to the previously published ADAMTS6 cDNA sequence, including a redefinition of the predicted active site motif. |
| Databáze: | OpenAIRE |
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