title: Purification of dFMR1-containing complexes using tandem affinity purification.
abstract: Fragile X syndrome results from the lack of FMR1 expression. To understand how the lack of FMR1 function leads to the syndrome, we are studying the Drosophila FMR1 related protein (dFMR1). We performed affinity purification of dFMR1-associated complexes from cultured Drosophila S2 cells and found that dFMR1 associates with a key component of RNA interference, AGO2. Our finding suggests cross talk between the fragile X syndrome protein and RNA interference. In this chapter, we describe a tandem affinity purification method to isolate the protein components and small RNAs in the dFMR1-associated complexes.
authors.affiliation: Department of Molecular Biology, Keio University School of Medicine, Tokyo, Japan.
authors.forename: Keita
authors.initials: K
authors.lastname: Miyoshi
authors.forename: Akiyo
authors.initials: A
authors.lastname: Ogino
authors.forename: Mikiko C
authors.initials: MC
authors.lastname: Siomi
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Siomi
mesh_terms: D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D005591:Chemical Fractionation / Q000379:methods*; D004330:Drosophila / Q000166:cytology; D029721:Drosophila Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; D051860:Fragile X Messenger Ribonucleoprotein 1; D034741:RNA, Small Interfering; C467328:FMR1 protein, Drosophila