A Genome-Wide Screen for Dendritically Localized RNAs Identifies Genes Required for Dendrite Morphogenesis.
Localizing messenger RNAs at specific subcellular sites is a conserved mechanism for targeting the synthesis of cytoplasmic proteins to distinct subcellular domains, thereby generating the asymmetric protein distributions necessary for cellular and developmental polarity. However, the full range of transcripts that are asymmetrically distributed in specialized cell types, and the significance of their localization, especially in the nervous system, are not known. We used the EP-MS2 method, which combines EP transposon insertion with the MS2/MCP in vivo fluorescent labeling system, to screen for novel localized transcripts in polarized cells, focusing on the highly branched Drosophila class IV dendritic arborization neurons. Of a total of 541 lines screened, we identified 55 EP-MS2 insertions producing transcripts that were enriched in neuronal processes, particularly in dendrites. The 47 genes identified by these insertions encode molecularly diverse proteins, and are enriched for genes that function in neuronal development and physiology. RNAi-mediated knockdown confirmed roles for many of the candidate genes in dendrite morphogenesis. We propose that the transport of mRNAs encoded by these genes into the dendrites allows their expression to be regulated on a local scale during the dynamic developmental processes of dendrite outgrowth, branching, and/or remodeling.
Department of Molecular Biology, Princeton University, NJ 08544.
Mala
M
Misra
Department of Molecular Biology, Princeton University, NJ 08544.
Hendia
H
Edmund
Department of Biochemistry, The University of Oxford, OX1 3QU, United Kingdom.
Darragh
D
Ennis
Department of Molecular Biology, Princeton University, NJ 08544.
Marissa A
MA
Schlueter
Department of Molecular Biology, Princeton University, NJ 08544.
Jessica E
JE
Marot
Department of Molecular Biology, Princeton University, NJ 08544.
Janet
J
Tambasco
Department of Molecular Biology, Princeton University, NJ 08544.
Ida
I
Barlow
European Molecular Biology Laboratory, 69117 Heidelberg, Germany.
Sara
S
Sigurbjornsdottir
European Molecular Biology Laboratory, 69117 Heidelberg, Germany.
Renjith
R
Mathew
Department of Biochemistry, The University of Oxford, OX1 3QU, United Kingdom.
Ana Maria
AM
Vallés
Biocenter, Institute of Developmental Biology, University of Cologne, 50674 Cologne, Germany.
Waldemar
W
Wojciech
Biocenter, Institute of Developmental Biology, University of Cologne, 50674 Cologne, Germany.
Siegfried
S
Roth
Department of Biochemistry, The University of Oxford, OX1 3QU, United Kingdom.
Ilan
I
Davis
European Molecular Biology Laboratory, 69117 Heidelberg, Germany Institute of Genetics, University of Cologne, 50674 Germany.
Maria
M
Leptin
Department of Molecular Biology, Princeton University, NJ 08544 gavis@princeton.edu.
Elizabeth R
ER
Gavis
D000818:Animals; D003712:Dendrites / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics / Q000254:growth & development; D018507:Gene Expression Regulation, Developmental; D049750:Genome, Insect; D055106:Genome-Wide Association Study; D007814:Larva / Q000235:genetics / Q000254:growth & development; D009024:Morphogenesis / Q000235:genetics*; D055495:Neurogenesis / Q000235:genetics*; D009474:Neurons / Q000378:metabolism; D017933:Peripheral Nervous System / Q000254:growth & development*; D012333:RNA, Messenger / Q000235:genetics*; D011984:Sensory Receptor Cells / Q000378:metabolism
D012333:RNA, Messenger
Drosophila peripheral nervous system; dendritic arborization neuron; local translation; mRNA localization; multidendritic neuron