title: RNA cargoes associating with FMRP reveal deficits in cellular functioning in Fmr1 null mice.
abstract: The Fragile X mental retardation-1 (Fmr1) gene encodes a multifunctional protein, FMRP, with intrinsic RNA binding activity. We have developed an approach, antibody-positioned RNA amplification (APRA), to identify the RNA cargoes associated with the in vivo configured FMRP messenger ribonucleoprotein (mRNP) complex. Using APRA as a primary screen, putative FMRP RNA cargoes were assayed for their ability to bind directly to FMRP using traditional methods of assessing RNA-protein interactions, including UV-crosslinking and filter binding assays. Approximately 60% of the APRA-defined mRNAs directly associate with FMRP. By examining a subset of these mRNAs and their encoded proteins in brain tissue from Fmr1 knockout mice, we have observed that some of these cargoes as well as the proteins they encode show discrete changes in abundance and/or differential subcellular distribution. These data are consistent with spatially selective regulation of multiple biological pathways by FMRP.
authors.affiliation: Department of Pharmacology, Department of Psychiatry, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA.
authors.forename: Kevin Y
authors.initials: KY
authors.lastname: Miyashiro
authors.forename: Andrea
authors.initials: A
authors.lastname: Beckel-Mitchener
authors.forename: T Patrick
authors.initials: TP
authors.lastname: Purk
authors.forename: Kevin G
authors.initials: KG
authors.lastname: Becker
authors.forename: Tanya
authors.initials: T
authors.lastname: Barret
authors.forename: Lei
authors.initials: L
authors.lastname: Liu
authors.forename: Salvatore
authors.initials: S
authors.lastname: Carbonetto
authors.forename: Ivan Jeanne
authors.initials: IJ
authors.lastname: Weiler
authors.forename: William T
authors.initials: WT
authors.lastname: Greenough
authors.forename: James
authors.initials: J
authors.lastname: Eberwine
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D015342:DNA Probes / Q000276:immunology; D051860:Fragile X Messenger Ribonucleoprotein 1; D005600:Fragile X Syndrome / Q000235:genetics / Q000378:metabolism*; D066298:In Vitro Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D011687:Purines / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins; D013347:Subcellular Fractions
chemical_list: D000911:Antibodies, Monoclonal; D015342:DNA Probes; D051860:Fragile X Messenger Ribonucleoprotein 1; D009419:Nerve Tissue Proteins; D011687:Purines; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C497684:Fmr1 protein, mouse