The third RNA recognition motif of Drosophila ELAV protein has a role in multimerization.

Nucleic Acids Research
Gakuta Toba, Kalpana White

Abstract

ELAV is a neuron-specific RNA-binding protein in Drosophila that is required for development and maintenance of neurons. ELAV regulates alternative splicing of Neuroglian and erect wing (ewg) transcripts, and has been shown to form a multimeric complex on the last ewg intron. The protein has three RNA recognition motifs (RRM1, 2 and 3) with a hinge region between RRM2 and 3. In this study, we used the yeast two-hybrid system to determine the multimerization domain of ELAV. Using deletion constructs, we mapped an interaction activity to a region containing most of RRM3. We found three conserved short sequences in RRM3 that were essential for the interaction, and also sufficient to give the interaction activity to RRM2 when introduced into it. In our in vivo functional assay, a mutation in one of the three sequences showed reduced activity in splicing regulation, underlining the functional importance of multimerization. However, RRM2 with the three RRM3 interaction sequences did not function as RRM3 in vivo, which suggested that multimerization is not the only function of RRM3. Our results are consistent with a model in which RRM3 serves as a bi-functional domain that interacts with both RNA and protein.

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Citations

Jun 27, 2008·Cellular and Molecular Life Sciences : CMLS·M N Hinman, H Lou
Dec 4, 2010·Briefings in Functional Genomics·Shivendra KishoreMihaela Zavolan
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Methods Mentioned

BETA
two-hybrid
PCR
PCRs
co-immunoprecipitation
immunoprecipitation
electrophoresis
transgenic
dissection
co-IP

Software Mentioned

Cn3D
ImageJ
ELAV

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