title: Identification of lncRNA MEG3 Binding Protein Using MS2-Tagged RNA Affinity Purification and Mass Spectrometry.
abstract: Long noncoding RNAs (lncRNAs) are nonprotein coding transcripts longer than 200 nucleotides. Recently in mammals, thousands of long noncoding RNAs have been identified and studied as key molecular players in different biological processes with protein complexes. As a long noncoding RNA, maternally expressed gene 3 (MEG3) plays an important role in many cellular processes. However, the mechanism underlying MEG3 regulatory effects remains enigmatic. By using the specific interaction between MS2 coat protein and MS2 RNA hairpin, we developed a method (MS2-tagged RNA affinity purification and mass spectrometry (MTRAP-MS)) to identify proteins that interact with MEG3. Mass spectrometry and gene ontology (GO) analysis showed that MEG3 binding proteins possess nucleotide binding properties and take part in transport, translation, and other biological processes. In addition, interleukin enhancer binding factor 3 (ILF3) and poly(A) binding protein, cytoplasmic 3 (PABPC3) were validated for their interaction with MEG3. These findings indicate that the newly developed method can effectively enrich lncRNA binding proteins and provides a strong basis for studying MEG3 functions.
authors.affiliation: Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, College of Life Science, Jilin University, Changchun, 130012, Jilin, People's Republic of China.
authors.forename: Shanshan
authors.initials: S
authors.lastname: Liu
authors.forename: Juanjuan
authors.initials: J
authors.lastname: Zhu
authors.forename: Taifeng
authors.initials: T
authors.lastname: Jiang
authors.forename: Yiran
authors.initials: Y
authors.lastname: Zhong
authors.forename: Yi
authors.initials: Y
authors.lastname: Tie
authors.forename: Yongge
authors.initials: Y
authors.lastname: Wu
authors.forename: Xiaofei
authors.initials: X
authors.lastname: Zheng
authors.forename: Yinghua
authors.initials: Y
authors.lastname: Jin
authors.forename: Hanjiang
authors.initials: H
authors.lastname: Fu
mesh_terms: D057809:HEK293 Cells; D006801:Humans; D013058:Mass Spectrometry; D050779:Nuclear Factor 90 Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D039102:Poly(A)-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D062085:RNA, Long Noncoding / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D050779:Nuclear Factor 90 Proteins; C467335:PABPC3 protein, human; D039102:Poly(A)-Binding Proteins; D062085:RNA, Long Noncoding; D016601:RNA-Binding Proteins