TY - JOUR
T1 - The third RNA recognition motif of Drosophila ELAV protein has a role in multimerization
AU - Toba, Gakuta
AU - White, Kalpana
N1 - Funding Information:
We thank M. Zhadina for help with the two-hybrid assay, P. Parmenter and E. Dougherty for technical assistance. We thank the Bloomington Drosophila Stock Center at Indiana University for providing fly stocks. The project described was supported by Grant Numbers P01NS044232 and P30 NS045713 from the National Institutes of Neurological Disorders and Stroke and Grant Number S10 RR16780 from the National Institute of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of Neurological Disorders and Stroke or the National Institute of Health.
PY - 2008/3
Y1 - 2008/3
N2 - 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.
AB - 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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U2 - 10.1093/nar/gkm1168
DO - 10.1093/nar/gkm1168
M3 - Article
C2 - 18203745
AN - SCOPUS:40249108471
SN - 0305-1048
VL - 36
SP - 1390
EP - 1399
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 4
ER -