Alternative polyadenylation in triple-negative breast tumors allows NRAS and c-JUN to bypass PUMILIO posttranscriptional regulation

Wayne O. Miles, Antonio Lembo, Angela Volorio, Elena Brachtel, Bin Tian, Dennis Sgroi, Paolo Provero, Nicholas Dyson

Research output: Contribution to journalArticle

Abstract

Alternative polyadenylation (APA) is a process that changes the posttranscriptional regulation and translation potential of mRNAs via addition or deletion of 3′ untranslated region (3′ UTR) sequences. To identify posttranscriptional-regulatory events affected by APA in breast tumors, tumor datasets were analyzed for recurrent APA events. Motif mapping of the changed 3′ UTR regions found that APA-mediated removal of Pumilio regulatory elements (PRE) was unusually common. Breast tumor subtype-specific APA profiling identified triple-negative breast tumors as having the highest levels of APA. To determine the frequency of these events, an independent cohort of triple-negative breast tumors and normal breast tissue was analyzed for APA. APA-mediated shortening of NRAS and c-JUN was seen frequently, and this correlated with changes in the expression of downstream targets. mRNA stability and luciferase assays demonstrated APA-dependent alterations in RNA and protein levels of affected candidate genes. Examination of clinical parameters of these tumors found those with APA of NRAS and c-JUN to be smaller and less proliferative, but more invasive than non-APA tumors. RT-PCR profiling identified elevated levels of polyadenylation factor CSTF3 in tumors with APA. Overexpression of CSTF3 was common in triple-negative breast cancer cell lines, and elevated CSTF3 levels were sufficient to induce APA of NRAS and c-JUN. Our results support the hypothesis that PRE-containing mRNAs are disproportionately affected by APA, primarily due to high sequence similarity in the motifs utilized by polyadenylation machinery and the PUM complex.
Original languageEnglish
Pages (from-to)7231 - 7241
Number of pages11
JournalCancer Research
Volume76
Issue number24
DOIs
Publication statusPublished - Dec 15 2016

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Polyadenylation
Breast Neoplasms
3' Untranslated Regions
mRNA Cleavage and Polyadenylation Factors
Neoplasms
Triple Negative Breast Neoplasms
RNA Stability
Protein Biosynthesis
Luciferases

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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Alternative polyadenylation in triple-negative breast tumors allows NRAS and c-JUN to bypass PUMILIO posttranscriptional regulation. / Miles, Wayne O.; Lembo, Antonio; Volorio, Angela; Brachtel, Elena; Tian, Bin; Sgroi, Dennis; Provero, Paolo; Dyson, Nicholas.

In: Cancer Research, Vol. 76, No. 24, 15.12.2016, p. 7231 - 7241.

Research output: Contribution to journalArticle

Miles, WO, Lembo, A, Volorio, A, Brachtel, E, Tian, B, Sgroi, D, Provero, P & Dyson, N 2016, 'Alternative polyadenylation in triple-negative breast tumors allows NRAS and c-JUN to bypass PUMILIO posttranscriptional regulation', Cancer Research, vol. 76, no. 24, pp. 7231 - 7241. https://doi.org/10.1158/0008-5472.CAN-16-0844
Miles, Wayne O. ; Lembo, Antonio ; Volorio, Angela ; Brachtel, Elena ; Tian, Bin ; Sgroi, Dennis ; Provero, Paolo ; Dyson, Nicholas. / Alternative polyadenylation in triple-negative breast tumors allows NRAS and c-JUN to bypass PUMILIO posttranscriptional regulation. In: Cancer Research. 2016 ; Vol. 76, No. 24. pp. 7231 - 7241.
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