NF90 selectively represses the translation of target mRNAs bearing an AU-rich signature motif.
The RNA-binding protein nuclear factor 90 (NF90) has been implicated in the stabilization, transport and translational control of several target mRNAs. However, a systematic analysis of NF90 target mRNAs has not been performed. Here, we use ribonucleoprotein immunoprecipitation analysis to identify a large subset of NF90-associated mRNAs. Comparison of the 3'-untranslated regions (UTRs) of these mRNAs led to the elucidation of a 25- to 30-nucleotide, RNA signature motif rich in adenines and uracils. Insertion of the AU-rich NF90 motif ('NF90m') in the 3'UTR of an EGFP heterologous reporter did not affect the steady-state level of the chimeric EGFP-NF90m mRNA or its cytosolic abundance. Instead, the translation of EGFP-NF90m mRNA was specifically repressed in an NF90-dependent manner, as determined by analysing nascent EGFP translation, the distribution of chimeric mRNAs on polysome gradients and the steady-state levels of expressed EGFP protein. The interaction of endogenous NF90 with target mRNAs was validated after testing both endogenous mRNAs and recombinant biotinylated transcripts containing NF90 motif hits. Further analysis showed that the stability of endogenous NF90 target mRNAs was not significantly influenced by NF90 abundance, while their translation increased when NF90 levels were reduced. In summary, we have identified an AU-rich RNA motif present in NF90 target mRNAs and have obtained evidence that NF90 represses the translation of this subset of mRNAs.
RNA Regulation Section, Laboratory of Cellular and Molecular Biology and Bioinformatics Unit, Research Resources Branch, National Institute on Aging-Intramural Research Program, National Institutes of Health, Baltimore, MD 21224, USA.
Yuki
Y
Kuwano
Rudolf
R
Pullmann
Bernard S
BS
Marasa
Kotb
K
Abdelmohsen
Eun Kyung
EK
Lee
Xiaoling
X
Yang
Jennifer L
JL
Martindale
Ming
M
Zhan
Myriam
M
Gorospe
D020413:3' Untranslated Regions; D000225:Adenine / Q000032:analysis; D006367:HeLa Cells; D006801:Humans; D050779:Nuclear Factor 90 Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D014176:Protein Biosynthesis; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D017423:Sequence Analysis, RNA; D014498:Uracil / Q000032:analysis
D020413:3' Untranslated Regions; D050779:Nuclear Factor 90 Proteins; D012333:RNA, Messenger; D014498:Uracil; D000225:Adenine