title: Monoclonal antibodies against Muscleblind-like 3, a protein with punctate nuclear localization.
abstract: Muscleblind-like 3 (MBNL3) belongs to a family of RNA binding proteins that regulate alternative splicing. We have generated a set of monoclonal antibodies (MAbs) against mouse MBNL3, three of which do not cross-react with the other muscleblind-like (MBNL) proteins, MBNL1 and MBNL2. Epitope mapping revealed that MAbs P1C7, P1E7, SP1C2, and P2E6 recognize distinct, non-overlapping segments of the MBNL3 polypeptide sequence. Immunohistochemical staining of proliferating muscle precursor cells localized MBNL3 to the nucleus in a punctate pattern, characteristic of subcellular structures in the nucleus enriched in pre-messenger RNA splicing factors. Although MBNL3 did not co-localize with SC35 and PSP1 (widely used markers of splicing speckles and paraspeckles), the punctate localization pattern of MBNL3 within interchromatin regions of the nucleus is highly predictive of proteins involved in pre-mRNA processing. Monoclonal antibodies specific for mouse MBNL3 will facilitate further investigation of the expression pattern and unique functions of this splicing factor during development and in different adult mouse tissues.
authors.affiliation: Department of Pharmacology, University of Washington, Seattle, Washington, USA.
authors.forename: Kyung-Soon
authors.initials: KS
authors.lastname: Lee
authors.forename: K A
authors.initials: KA
authors.lastname: Lewis
authors.forename: Susan
authors.initials: S
authors.lastname: Tom
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Wayner
authors.forename: Edith H
authors.initials: EH
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification; D000918:Antibody Specificity; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D018604:Epitope Mapping; D004926:Escherichia coli; D018507:Gene Expression Regulation, Developmental; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D007114:Immunization; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism*; D020033:Protein Isoforms / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D012322:RNA Precursors / Q000378:metabolism; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D002843:Chromatin; C522920:MBNL3 protein, mouse; D020033:Protein Isoforms; D012322:RNA Precursors; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins