TY - JOUR
T1 - Dispatched Homolog 2 is targeted by miR-214 through a combination of three weak microRNA recognition sites
AU - Li, Nan
AU - Flynt, Alex S.
AU - Kim, H. Rosemary
AU - Solnica-krezel, Lilianna
AU - Patton, James G.
N1 - Funding Information:
This work was supported by NIH grant GM 075790 to J.G.P. A.F. was supported in part by T32 GM 008554. Funding to pay the Open Access publication charges for this article was provided by NIH GM 075790.
PY - 2008/8
Y1 - 2008/8
N2 - MicroRNAs (miRNAs) regulate gene expression by inhibiting translation of target mRNAs through pairing with miRNA recognition elements (MREs), usually in 3′ UTRs. Because pairing is imperfect, identification of bona fide mRNA targets presents a challenge. Most target recognition algorithms strongly emphasize pairing between nucleotides 2-8 of the miRNA (the 'seed' sequence) and the mRNA but adjacent sequences and the local context of the 3′ UTR also affect targeting. Here, we show that dispatched 2 is a target of miR-214. In zebrafish, dispatched 2 is expressed in the telencephalon and ventral hindbrain and is essential for normal zebrafish development. Regulation of dispatched 2 by miR-214 is via pairing with three, noncanonical, weak MREs. By comparing the repression capacity of GFP reporters containing different dispatched 2 sequences, we found that a combination of weak sites, which lack canonical seed pairing, can effectively target an mRNA for silencing. This finding underscores the challenge that prediction algorithms face and emphasizes the need to experimentally validate predicted MREs.
AB - MicroRNAs (miRNAs) regulate gene expression by inhibiting translation of target mRNAs through pairing with miRNA recognition elements (MREs), usually in 3′ UTRs. Because pairing is imperfect, identification of bona fide mRNA targets presents a challenge. Most target recognition algorithms strongly emphasize pairing between nucleotides 2-8 of the miRNA (the 'seed' sequence) and the mRNA but adjacent sequences and the local context of the 3′ UTR also affect targeting. Here, we show that dispatched 2 is a target of miR-214. In zebrafish, dispatched 2 is expressed in the telencephalon and ventral hindbrain and is essential for normal zebrafish development. Regulation of dispatched 2 by miR-214 is via pairing with three, noncanonical, weak MREs. By comparing the repression capacity of GFP reporters containing different dispatched 2 sequences, we found that a combination of weak sites, which lack canonical seed pairing, can effectively target an mRNA for silencing. This finding underscores the challenge that prediction algorithms face and emphasizes the need to experimentally validate predicted MREs.
UR - http://www.scopus.com/inward/record.url?scp=48349136544&partnerID=8YFLogxK
U2 - 10.1093/nar/gkn388
DO - 10.1093/nar/gkn388
M3 - Article
C2 - 18583362
AN - SCOPUS:48349136544
SN - 0305-1048
VL - 36
SP - 4277
EP - 4285
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 13
ER -