Role of miR-103 and miR-107 in Regulation of CDK5R1 Expression and in Cellular Migration

被引:84
作者
Moncini, Silvia [1 ]
Salvi, Alessandro [2 ]
Zuccotti, Paola [1 ]
Viero, Gabriella [3 ,4 ]
Quattrone, Alessandro [3 ]
Barlati, Sergio [2 ]
De Petro, Giuseppina [2 ]
Venturin, Marco [1 ]
Riva, Paola [1 ]
机构
[1] Univ Milan, Dept Biol & Genet Med Sci, Milan, Italy
[2] Univ Brescia, Dept Biomed Sci & Biotechnol, Div Biol & Genet, Brescia, Italy
[3] Univ Trento, CIBIO, Trento, Italy
[4] CNR, Inst Biophys, Trento, Italy
来源
PLOS ONE | 2011年 / 6卷 / 05期
关键词
CYCLIN-DEPENDENT KINASE-5; NEURONAL-SPECIFIC ACTIVATOR; MICRORNA EXPRESSION; NEURITE OUTGROWTH; MESSENGER-RNA; TAU-PROTEIN; MOUSE MODEL; GENES; P35; PRECURSOR;
D O I
10.1371/journal.pone.0020038
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
CDK5R1 encodes p35, a specific activator of the serine/threonine kinase CDK5, which plays crucial roles in CNS development and maintenance. CDK5 activity strongly depends on p35 levels and p35/CDK5 misregulation is deleterious for correct CNS function, suggesting that a tightly controlled regulation of CDK5R1 expression is needed for proper CDK5 activity. Accordingly, CDK5R1 expression was demonstrated to be controlled at both transcriptional and post-transcriptional levels, but a possible regulation through microRNAs (miRNAs) has never been investigated. We predicted, within the large CDK5R1 3'UTR several miRNA target sites. Among them, we selected for functional studies miR-103 and miR-107, whose expression has shown a strong inverse correlation with p35 levels in different cell lines. A significant reduction of CDK5R1 mRNA and p35 levels was observed after transfection of SK-N-BE neuroblastoma cells with the miR-103 or miR-107 precursor (pre-miR-103 or pre-miR-107). Conversely, p35 levels significantly increased following transfection of the corresponding antagonists (anti-miR-103 or anti-miR-107). Moreover, the level of CDK5R1 transcript shifts from the polysomal to the subpolysomal mRNA fraction after transfection with pre-miR-107 and, conversely, from the subpolysomal to the polysolmal mRNA fraction after transfection with anti-miR-107, suggesting a direct action on translation efficiency. We demonstrate, by means of luciferase assays, that miR-103 and miR-107 are able to directly interact with the CDK5R1 3'-UTR, in correspondence of a specific target site. Finally, miR-103 and miR-107 overexpression, as well as CDK5R1 silencing, caused a reduction in SK-N-BE migration ability, indicating that these miRNAs affect neuronal migration by modulating CDK5R1 expression. These findings indicate that miR-103 and miR-107 regulate CDK5R1 expression, allowing us to hypothesize that a miRNA-mediated mechanism may influence CDK5 activity and the associated molecular pathways.
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页数:13
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