• BMB Reports
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• v.47 no.11
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• pp.643-648
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• 2014

Stanniocalcin (STC), a glycoprotein hormone originally discovered in fish, has been implicated in calcium and phosphate homeostasis. While fishes and mammals possess two STC homologs (STC1 and STC2), the physiological roles of STC2 are largely unknown compared with those of STC1. In this study, we identified Ran-binding protein M (RanBPM) as a novel binding partner of STC2 using yeast two-hybrid screening. The interaction between STC2 and RanBPM was confirmed in mammalian cells by immunoprecipitation. STC2 enhanced the RanBPM-mediated transactivation of liganded androgen receptor (AR), but not thyroid receptor ${\beta}$, glucocorticoid receptor, or estrogen receptor ${\beta}$. We also found that AR interacted with RanBPM in both the absence and presence of testosterone (T). Furthermore, we discovered that STC2 recruits RanBPM/AR complex in T-dependent manner. Taken together, our findings suggest that STC2 is a novel RanBPM-interacting protein that promotes AR transactivation.

• Biomolecules & Therapeutics
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• v.17 no.1
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• pp.12-16
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• 2009

One of the WHSC1/MMSET/NSD2 variant RE-IIBP is a histone H3-K27 methyltransferase with transcriptional repression activity. Overexpression of RE-IIBP in various types of leukemia suggests it's role in leukemogenesis. Here we identify two proteins, KRAB zinc finger protein ZNF 350 and enolase-1 as RE-IIBP interacting proteins by yeast two-hybrid screening and confirmed direct interaction in vivo and in vitro. Both proteins have been known for their role in transcriptional repression. Reporter assays using transient transfection demonstrated that both ZNF 350 and enolase-1 proteins synergistically repressed transcription with RE-IIBP, respectively. These results indicate both proteins have roles in RE-IIBP mediated transcriptional repression by involving co-repressor complex.

• Korean Journal of Microbiology
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• v.50 no.2
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• pp.95-100
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• 2014

Hrq1 is a novel member of RecQ helicase family, found in fungal genomes by bioinformatics analyses. It is most homologous to human RECQL4 and recent genetic and biochemical studies suggested that it may play roles in the maintenance of genome stability. In this study, we investigated yeast two-hybrid interactions between Hrq1 and the yeast genes homologous to the human genes that are known to interact with RECQL4. Among the 11 genes tested, Rad14, a nucleotide excision repair (NER) factor, was found to interact with Hrq1. In addition, pull-down assay with the purified proteins revealed direct protein-protein interaction between Hrq1 and Rad14. The yeast two-hybrid interaction was enhanced by the DNA damage induced by 4-nitroquinoline-1-oxide, which was dependent on the presence of Rad4, a key NER factor. These results suggest that Hrq1 may function in NER through interaction with Rad14.

• Journal of Microbiology and Biotechnology
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• v.22 no.3
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• pp.378-384
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• 2012

In aquatic invertebrates, particularly marine gastropods, organotin compounds induce irreversible sexual abnormality in females, which is termed imposex, at very low concentrations. Organotin compounds are agonists for nuclear receptors such as RXRs and $PPAR{\gamma}$. However, the imposex phenomenon has not been reported to act as an antagonist on estrogen receptors in other species, including vertebrates and invertebrates. In order to gain insights into the antagonistic activity of organotin compounds on estrogen receptors (ERs), we examined the inhibitive effect of these compounds on estradiol-dependent ${\beta}$-galactosidase activity using the yeast two-hybrid detection system consisting of a combination of the human estrogen receptor ($hER{\beta}$) ligand-binding domain and the co-activator steroid receptor co-activator-1 (SRC1). Tributyltin-hydroxide (TBT-OH) and triphenyltin-chlorine (TPT-Cl) exhibited an inhibitive effect on $E_2$-dependent transcriptional activity, similar to antagonistic chemicals such as 4-hydroxytamoxifen (OHT) or ICI 182,780, at a very low concentration of $10^{-14}$ M TBT or $10^{-10}$ M TPT, respectively. The yeast growth and transcriptional activity with transcriptional factor GAL4 did not exhibit any effect at the tested concentration of TBT or TPT. Moreover, the yeast two-hybrid system using the interaction between p53 and the T antigen of SV40 large did not describe any effect at the tested concentration of OHT or ICI 182,780. However, the interaction between p53 and T antigen was inhibited at a TBT or TPT concentration of $10^{-9}$ M, respectively. These results indicate that TBT and TPT act as inhibitors of ER-dependent reporter gene transcriptional activation and of the interaction between $hER{\beta}$ LBD and the co-activator SRC1 in the yeast two-hybrid system. Consequently, our data could partly explain the occurrence of organotin compound-induced imposex on the endocrine system of mammals, including humans.

• Korean Journal of Microbiology
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• v.53 no.1
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• pp.49-54
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• 2017

Sus1 / ENY2 is a tiny conserved protein that is involved in chromatin remodeling and mRNA biogenesis. Sus1 is associated to two nuclear complexes, the transcriptional coactivator SAGA and the nuclear pore associated TREX2. In fission yeast, Schizosaccharomyces pombe, ortholog of Sus1 / ENY2 was identified from the genome database. Tetrad analysis showed that the S. pombe sus1 is not essential for growth. However, deletion of the sus1 gene caused cold-sensitive growth retardation with slight accumulation of $poly(A)^+$ RNA in the nucleus. And the Sus1-GFP protein is localized mainly in the nucleus. Yeast two-hybrid analysis and co-immunoprecipitation experiment showed that Sus1 interacts with Sac3, another subunit of TREX2 complex. These results suggest that S. pombe Sus1 is also involved in mRNA export from the nucleus as a component of TREX-2 complex.

• BMB Reports
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• v.34 no.1
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• pp.85-89
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• 2001

Both topoisomerase $II{\alpha}$ and $II{\beta}$ east as phosphoproteins in the cells. Recently it was reported that DNA topoisomerase $II{\alpha}$ associates with and is phosphorylated by the extracellular signal-regulated kinase 2 (ERK2). Also, ERK2 stimulates the activity of topoisomerase II by a phosphorylation-independent manner [Shapiro et al., (1999) Mol. Cell. Biol. 19, 3551-3560]. In this study, a yeast two-hybrid system was used to investigate the binding site between topoisomerase $II{\alpha}$ or $II{\beta}$ and ERK2. The two-hybrid test clearly showed that topoisomerase $II{\beta}$ residues 1099-1263, and topoisomerase $II{\alpha}$ residues 1078-1182, mediate the interaction with ERK2, and that the leucine zipper motifs of topoisomerase $II{\alpha}$ and $II{\beta}$ are not required for its physical binding to ERK2. Our results suggest that topoisomerase $II{\beta}$ residues 1099-1263, and topoisomerase $II{\alpha}$ residues 1078-1182, may be common binding sites for activator proteins.

• Korean Journal of Microbiology
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• v.54 no.4
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• pp.325-329
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• 2018

Thp1/PCID2, PCI domain-containing protein, is a component of the evolutionally conserved TREX-2 complex linking mRNA transcription and export. In fission yeast, Schizosaccharomyces pombe, the pci2 (SPBC1105.07c) gene encodes a PCI domain-containing protein that is predicted as a fission yeast orthologue of Thp1 (in budding yeast)/PCID2 (in human). Repression of pci2 expression inhibited both growth and mRNA export. And over-expression of pci2 also exhibited growth retardation with slight accumulation of $poly(A)^+$ RNA in the nucleus. Moreover, yeast two-hybrid and co-immunoprecipitation analysis showed that the Pci2 protein physically interacted with Sac3 and Dss1, which are members of TREX-2 complex. These observations support that the S. pombe Pci2 protein, as a component of TREX-2 complex, is implicated in mRNA export.

• Biomedical Science Letters
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• v.13 no.2
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• pp.127-133
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• 2007

Heterotrimeric GTP binding proteins (G proteins) mediate signal generated by neurotransmitter and hormones. Among all G proteins, Go is the most abundant in brain but its role in brain is not clearly understood. To determine the function of the alpha subunit of Go (Go$\alpha$), we search for the interacting partner of Go$\alpha$ in brain using yeast two-hybrid system. A resistant to inhibitor of cholinesterase (Ric-8B) was identified as a Go$\alpha$ interacting protein. We confirmed interaction between Go$\alpha$ and Ric-8b employing in vitro affinity binding assay and showed that the Ric-8b increased the function of Go$\alpha$. Our findings indicate that Ric-8b is possible guanine nucleotide exchange factor for Go$\alpha$.

• BMB Reports
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• v.37 no.1
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• pp.45-52
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• 2004

The goal of interaction proteomics that studies the protein-protein interactions of all expressed proteins is to understand biological processes that are strictly regulated by these interactions. The availability of entire genome sequences of many organisms and high-throughput analysis tools has led scientists to study the entire proteome (Pandey and Mann, 2000). There are various high-throughput methods for detecting protein interactions such as yeast two-hybrid approach and mass spectrometry to produce vast amounts of data that can be utilized to decipher protein functions in complicated biological networks. In this review, we discuss recent developments in analytical methods for large-scale protein interactions and the future direction of interaction proteomics.

• Biomedical Science Letters
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• v.13 no.4
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• pp.305-311
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• 2007

cAMP-dependent protein kinase A (PKA) is the best-characterized protein kinases and has served as a model of the structure and regulation of cAMP-binding protein as well as of protein kinases. To determine the function of PKA in development, we employed the yeast two-hybrid system to screen for catalytic subunit of PKA $(C\alpha)$ interacting partners in a cDNA library from mouse embryo. A Testis-brain RNA-binding protein (TB-RBP), specifically bound to $C\alpha$. This interaction was verified by several biochemical analysis. Our findings indicate that $C\alpha$ can modulate nucleic acid binding proteins of TB-RBP and provide insights into the diverse role of PKA.