Role of Pur alpha in targeting mRNA to sites of translation in hippocampal neuronal dendrites.
Using genetic inactivation in the mouse, PURA, encoding Pur alpha, is demonstrated to be essential for developmentally-timed dendrite formation in the cerebellum and hippocampus. Comparison of RNA species bound by Pur alpha prompts the hypothesis that Pur alpha functions with non-coding RNA in transport of certain mRNA molecules to sites of translation in dendrites. Pur alpha binds to human BC200 RNA, implicated in dendritic targeting, and this has homologies to 7SL RNA, implicated in compartmentalized translation. Results using hippocampal rat neurons in situ show that Pur alpha binds to BC1 RNA, implicated in dendritic targeting as a mouse counterpart of BC200, and to mRNA molecules translated in dendrites; Pur alpha is specifically located in dendrites, where it is colocalized with Map2, but not in axons, where it fails to colocalize with Ankyrin G. Pur alpha and Staufen are colocalized at dendritic sites of mRNA translation. Microtubule disruptors inhibit Pur alpha dendritic targeting and allow its mislocalization to axons. Using mouse brain, double-RNA immunoprecipitation places Pur alpha together with Staufen or FMRP on BC1 RNA and specific mRNA species in vivo. These results help define a mechanism by which Pur alpha targets specific mRNA molecules to sites of dendritic translation.
Johnson
Edward M
EM
Department of Pathology, Mount Sinai School of Medicine, New York, New York, USA. johnsoem@evms.edu
Kinoshita
Yayoi
Y
Weinreb
David B
DB
Wortman
Margaret J
MJ
Simon
Ruth
R
Khalili
Kamel
K
Winckler
Bettina
B
Gordon
Jennifer
J
D000818:Animals; D000831:Animals, Newborn; D017487:Ankyrins / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D002531:Cerebellum / Q000254:growth & development / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000172:deficiency / Q000502:physiology*; D003712:Dendrites / Q000502:physiology*; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods; D004622:Embryo, Mammalian; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology* / Q000254:growth & development / Q000378:metabolism; D007150:Immunohistochemistry / Q000379:methods; D047468:Immunoprecipitation / Q000379:methods; D051379:Mice; D018345:Mice, Knockout; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000172:deficiency; D009474:Neurons / Q000166:cytology*; D015739:Nocodazole / Q000494:pharmacology; D014176:Protein Biosynthesis / Q000187:drug effects / Q000502:physiology*; D062085:RNA, Long Noncoding; D012333:RNA, Messenger / Q000378:metabolism; D022661:RNA, Untranslated; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D020736:Ribonucleoproteins, Small Cytoplasmic / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014157:Transcription Factors / Q000502:physiology*
D017487:Ankyrins; D004268:DNA-Binding Proteins; D051860:Fragile X Messenger Ribonucleoprotein 1; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; D015739:Nocodazole; D062085:RNA, Long Noncoding; D012333:RNA, Messenger; D022661:RNA, Untranslated; D016601:RNA-Binding Proteins; D020736:Ribonucleoproteins, Small Cytoplasmic; D014157:Transcription Factors; C490614:Ank3 protein, rat; C489663:Bc1 long-non-coding RNA, rat; C489787:MAP2 protein, rat; C508559:PurA protein, rat; C121485:Pura protein, mouse; C492186:Stau1 protein, rat