| Title |
Functional mapping of the plant small RNA methyltransferase: HEN1 physically interacts with HYL1 and DICER-LIKE 1 proteins / |
| Authors |
Baranauskė, Simona ; Mickutė, Milda ; Plotnikova, Aleksandra ; Finke, Andreas ; Venclovas, Česlovas ; Klimašauskas, Saulius ; Vilkaitis, Giedrius |
| DOI |
10.1093/nar/gkv102 |
| Full Text |
|
| Is Part of |
Nucleic acids research.. Oxford : Oxford University Press. 2015, Vol. 43, No. 5, p. 2802-2812.. ISSN 0305-1048. eISSN 1362-4962 |
| Keywords [eng] |
Methylation ; miRNA biogenesis ; 2'-O-methyltransferase HEN1 |
| Abstract [eng] |
Methylation of 3'-terminal nucleotides of miRNA/miRNA* is part of miRNAs biogenesis in plants but is not found in animals. In Arabidopsis thaliana this reaction is carried out by a multidomain AdoMet-dependent 2'-O-methyltransferase HEN1. Using deletion and structure-guidedmutational analysis, we show that the double-stranded RNA-binding domains R1 and R2 of HEN1 make significant but uneven contributions to substrate RNA binding, and map residues in each domain responsible for this function. Using GST pull-down assays and yeast two-hybrid analysis we demonstrate direct HEN1 interactions, mediated by its FK506-binding protein-like domain and R2 domain, with the microRNA biogenesis protein HYL1. Furthermore, we find that HEN1 forms a complex with DICER-LIKE 1 (DCL1) ribonuclease, another key protein involved in miRNA biogenesis machinery. In contrast, no direct interaction is detectable between HEN1 and SERRATE. On the basis of these findings, we propose a mechanism of plant miRNA maturation which involves binding of the HEN1 methyltransferase to the DCL1•HYL1•miRNA complex excluding the SERRATE protein. |
| Published |
Oxford : Oxford University Press |
| Type |
Journal article |
| Language |
English |
| Publication date |
2015 |
| CC license |
|
