Fragile X protein family member FXR1P is regulated by microRNAs

Anne Cheever, Ernest Blackwell, Stephanie Ceman

Research output: Contribution to journalArticle

Abstract

FXR1P is one of two autosomal paralogs of the fragile X mental retardation protein FMRP. The absence of FMRP causes fragile X syndrome, the leading cause of hereditary mental retardation. FXR1P plays an important role in normal muscle development and has been implicated in facioscapulohumeral muscular dystrophy (FSHD). Its absence also causes cardiac abnormalities in both mice and zebrafish. To examine miRNA-mediated regulation of FMRP and FXR1P, we studied their expression in a conditional Dicer knockdown cell line, DT40. We found that FXR1P, but not FMRP, is significantly increased upon Dicer knockdown and the consequent reduction of miRNAs, suggesting that FXR1P is regulated by miRNAs while FMRP is not in DT40 cells. Expression of a luciferase reporter bearing the 3′ untranslated region (3′UTR) of FXR1 was significantly increased in the absence of miRNAs, confirming miRNA-mediated regulation of FXR1P, while a luciferase reporter bearing the FMR1 3′UTR was not. We identified one of the regulatory regions in the 3′UTR of FXR1 by removing a conserved, 8-nucleotide miRNA seed sequence common to miRNAs 25, 32, 92, 363, and 367 and demonstrated loss of miRNA-mediated suppression. Treatment with specific miRNA hairpin inhibitors to each of the miRNAs in the seed sequence showed that miRs 92b, 363, and 367 regulated FXR1P expression. Accordingly, overexpression of the miRNA 367 mimic significantly decreased endogenous FXR1P expression in human cell lines HEK-293T and HeLa. We report for the first time that FXR1P is regulated through miRNA binding, with one site being the miR-25/32/92/363/367 seed sequence.

Original languageEnglish (US)
Pages (from-to)1530-1539
Number of pages10
JournalRNA
Volume16
Issue number8
DOIs
StatePublished - Aug 1 2010

Fingerprint

MicroRNAs
Proteins
3' Untranslated Regions
Seeds
Luciferases
Fragile X Mental Retardation Protein
Facioscapulohumeral Muscular Dystrophy
Fragile X Syndrome
Cell Line
Muscle Development
Nucleic Acid Regulatory Sequences
Zebrafish
Intellectual Disability
Nucleotides

Keywords

  • FMRP
  • FXR1P
  • Translation regulation
  • miRNA

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Fragile X protein family member FXR1P is regulated by microRNAs. / Cheever, Anne; Blackwell, Ernest; Ceman, Stephanie.

In: RNA, Vol. 16, No. 8, 01.08.2010, p. 1530-1539.

Research output: Contribution to journalArticle

Cheever, Anne ; Blackwell, Ernest ; Ceman, Stephanie. / Fragile X protein family member FXR1P is regulated by microRNAs. In: RNA. 2010 ; Vol. 16, No. 8. pp. 1530-1539.
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