Cap-independent translation through the p27 5′-UTR

Hong Jiang, Jennifer Coleman, Robin Miskimins, Rekha Srinivasan, W. Keith Miskimins

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Several recent publications have explored cap-independent translation through an internal ribosome entry site (IRES) in the 5′-UTR of the mRNA encoding the cyclin-dependent kinase inhibitor p27. The major experimental tool used in these reports was the use of bicistronic reporter constructs in which the 5′-UTR was inserted between the upstream and downstream cistrons. None of these reports has completely ruled out the possibility that the 5′-UTR has either cryptic promoter activity or a cryptic splice acceptor site. Either of these possibilities could result in expression of a monocistronic mRNA encoding the downstream cistron and false identification of an IRES. Indeed, Liu et al. recently published data suggesting that the p27 5′-UTR harbors cryptic promoter activity which accounts for its putative IRES activity. In this report, we have explored this potential problem further using promoterless bicistronic constructs coupled with RNase protection assays, siRNA knockdown of individual cistrons, RT-PCR to detect mRNA encoded by the bicistronic reporter with high sensitivity, direct transfection of bicistronic mRNAs, and insertion of an iron response element into the bicistronic reporter. The results do not support the conclusion that the p27 5′-UTR has significant functional promoter activity or cryptic splice sites, but rather that it is able to support cap-independent initiation of translation.

Original languageEnglish (US)
Pages (from-to)4767-4778
Number of pages12
JournalNucleic acids research
Volume35
Issue number14
DOIs
StatePublished - Jul 1 2007

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5' Untranslated Regions
RNA Splice Sites
Messenger RNA
Cyclin-Dependent Kinase Inhibitor p27
Genes
Response Elements
Ribonucleases
Small Interfering RNA
Transfection
Iron
Polymerase Chain Reaction
Internal Ribosome Entry Sites

ASJC Scopus subject areas

  • Genetics

Cite this

Jiang, H., Coleman, J., Miskimins, R., Srinivasan, R., & Miskimins, W. K. (2007). Cap-independent translation through the p27 5′-UTR. Nucleic acids research, 35(14), 4767-4778. https://doi.org/10.1093/nar/gkm512

Cap-independent translation through the p27 5′-UTR. / Jiang, Hong; Coleman, Jennifer; Miskimins, Robin; Srinivasan, Rekha; Miskimins, W. Keith.

In: Nucleic acids research, Vol. 35, No. 14, 01.07.2007, p. 4767-4778.

Research output: Contribution to journalArticle

Jiang, H, Coleman, J, Miskimins, R, Srinivasan, R & Miskimins, WK 2007, 'Cap-independent translation through the p27 5′-UTR', Nucleic acids research, vol. 35, no. 14, pp. 4767-4778. https://doi.org/10.1093/nar/gkm512
Jiang H, Coleman J, Miskimins R, Srinivasan R, Miskimins WK. Cap-independent translation through the p27 5′-UTR. Nucleic acids research. 2007 Jul 1;35(14):4767-4778. https://doi.org/10.1093/nar/gkm512
Jiang, Hong ; Coleman, Jennifer ; Miskimins, Robin ; Srinivasan, Rekha ; Miskimins, W. Keith. / Cap-independent translation through the p27 5′-UTR. In: Nucleic acids research. 2007 ; Vol. 35, No. 14. pp. 4767-4778.
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