The role of PRL1 in siRNA biogenesis.

Abstract

<p>(A) PRL1 interacts with DCL3 and DCL4. Co-IP was performed to detect the interaction of PRL1 with DCL3 or DCL4. MBP and MBP-PRL1 fused protein were expressed in <i>E.coli</i>. YFP, DCL3-YFP and DCL4-YFP were expressed in <i>N. benthamiana</i> leaves. Anti-YFP was used for IP. For loading, ten percent and one percent of input proteins were used for IP and Co-IP, respectively. (B) <i>prl1-2</i> impairs siRNA production from double-stranded RNAs (dsRNAs). Protein extracts isolated from inflorescences of Col, <i>prl1-2</i> and <i>prl1-2</i> containing a PRL1-YFP transgene were incubated dsRNAs for 120 min. dsRNAs were synthesized through <i>in vitro</i> transcription of a DNA fragment (5′ portion of <i>UBQ5</i> gene, ∼460 bp) under the presence of [α-³²P] UTP.</p