1. Identification of Binding Proteins for TSC22D1 Family Proteins Using Mass Spectrometry.
- Author
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Kamimura R, Uchida D, Kanno SI, Shiraishi R, Hyodo T, Sawatani Y, Shimura M, Hasegawa T, Tsubura-Okubo M, Yaguchi E, Komiyama Y, Fukumoto C, Izumi S, Fujita A, Wakui T, and Kawamata H
- Subjects
- Alternative Splicing, Cell Differentiation, Cell Line, Cell Nucleus metabolism, Cytoplasm metabolism, DNA Damage, HEK293 Cells, Humans, Mass Spectrometry, Mitochondria metabolism, Protein Binding, Protein Interaction Maps, GTP-Binding Proteins metabolism, Histones metabolism, Nuclear Proteins metabolism, Repressor Proteins metabolism
- Abstract
TSC-22 (TGF-β stimulated clone-22) has been reported to induce differentiation, growth inhibition, and apoptosis in various cells. TSC-22 is a member of a family in which many proteins are produced from four different family genes. TSC-22 (corresponding to TSC22D1-2) is composed of 144 amino acids translated from a short variant mRNA of the TSC22D1 gene. In this study, we attempted to determine the intracellular localizations of the TSC22D1 family proteins (TSC22D1-1, TSC-22 (TSC22D1-2), and TSC22(86) (TSC22D1-3)) and identify the binding proteins for TSC22D1 family proteins by mass spectrometry. We determined that TSC22D1-1 was mostly localized in the nucleus, TSC-22 (TSC22D1-2) was localized in the cytoplasm, mainly in the mitochondria and translocated from the cytoplasm to the nucleus after DNA damage, and TSC22(86) (TSC22D1-3) was localized in both the cytoplasm and nucleus. We identified multiple candidates of binding proteins for TSC22D1 family proteins in in vitro pull-down assays and in vivo binding assays. Histone H1 bound to TSC-22 (TSC22D1-2) or TSC22(86) (TSC22D1-3) in the nucleus. Guanine nucleotide-binding protein-like 3 (GNL3), which is also known as nucleostemin, bound to TSC-22 (TSC22D1-2) in the nucleus. Further investigation of the interaction of the candidate binding proteins with TSC22D1 family proteins would clarify the biological roles of TSC22D1 family proteins in several cell systems.
- Published
- 2021
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