title: Characterization of plant polyadenylation complexes by using tandem affinity purification.
abstract: Messenger RNA in eukaryotic cells is initially produced as a nascent transcript (pre-mRNA) without a polyadenine [poly(A)] tail to the 3' end. The precise cleavage of the pre-mRNA and addition of a poly(A) track need the communication between cis-elements in the pre-mRNA sequences and transacting protein factors recognizing them. Based on homology analyses, Arabidopsis cleavage and polyadenylation specificity factor (AtCPSF) complex should play a critical role in pre-mRNA 3' end processing. Here we describe the isolation of AtCPSF complex by using a tandem affinity purification (TAP) method. We demonstrate that TAP is a potent protein complex isolating approach that can fulfill a downstream protein identification purpose based on mass spectrometry techniques.
authors.affiliation: Department of Plant Pathology, College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China, hzhao@njau.edu.cn.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Zhao
authors.forename: Xinfu
authors.initials: X
authors.lastname: Ye
authors.forename: Qingshun Quinn
authors.initials: QQ
authors.lastname: Li
mesh_terms: D017360:Arabidopsis / Q000737:chemistry*; D029681:Arabidopsis Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D039223:Cleavage And Polyadenylation Specificity Factor / Q000737:chemistry / Q000302:isolation & purification*; D013058:Mass Spectrometry; D026723:Polyadenylation
chemical_list: D029681:Arabidopsis Proteins; C514052:CPSF73-I protein, Arabidopsis; D039223:Cleavage And Polyadenylation Specificity Factor