Molecular Shape and ATP Binding Activity of Rat p50, a Putative Mammalian Homologue of RuvB DNA Helicase

Kikuchi, Noriko; Gohshi, Takashi; Kawahire, Shigeru; Tachibana, Taro; Yoneda, Yoshihiro; Isobe, Toshiaki; Lim, Chun Ren; Kohno, Kenji; Ichimura, Tsuyoshi; Omata, Saburo; Horigome, Tsuneyoshi

doi:10.1093/oxfordjournals.jbchem.a022312

Cited by 11 publications

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“…Because TIH1 had been shown to be an essential gene (9,19,22), we first asked if TIH2 is also required for viability. The TIH2 gene was disrupted as described under "Experimental Procedures."…”

Section: Tih2mentioning

confidence: 99%

“…The Walker A Domain of Tih2p Is Essential for Its Function-The only recognizable functional motifs in Tih2p are the so-called Walker A and B motifs, which are involved in ATP binding/hydrolysis and are the region of Tih2p most homologous to the prokaryotic helicase, RuvB (9,10,35). To investigate whether the helicase domain of Tih2p contributes to its essential physiological role, the invariant lysine at position 81 in the major GX 4 GKT nucleotide-binding loop was changed to arginine (K81R) by site-directed mutagenesis (Fig.…”

Section: Tih2mentioning

confidence: 99%

“…Several O-GlcNAc-containing proteins such as nucleoporins as well as nonglycosylated proteins like importin ␤ were isolated (9). Two RuvB-like proteins, p50 (9) and p47 (10), were also isolated by this method probably because of their interaction with O-GlcNAc-bearing proteins.…”

mentioning

confidence: 99%

“…Two RuvB-like proteins, p50 (9) and p47 (10), were also isolated by this method probably because of their interaction with O-GlcNAc-bearing proteins. RuvB is a prokaryotic DNA helicase, and its helicase activity and DNA binding affinity are enhanced by interaction with RuvA (11,12).…”

mentioning

confidence: 99%

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The Saccharomyces cerevisiae RuvB-like Protein, Tih2p, Is Required for Cell Cycle Progression and RNA Polymerase II-directed Transcription

Lim¹,

Kimata²,

Ohdate³

et al. 2000

Journal of Biological Chemistry

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Two highly conserved RuvB-like putative DNA helicases, p47/TIP49b and p50/TIP49a, have been identified in the eukaryotes. Here, we study the function of Saccharomyces cerevisiae TIH2, which corresponds to mammalian p47/TIP49b. Tih2p is required for vegetative cell growth and localizes in the nucleus. Immunoprecipitation analysis revealed that Tih2p tightly interacts with Tih1p, the counterpart of mammalian p50/TIP49a, which has been shown to interact with the TATA-binding protein and the RNA polymerase II holoenzyme complex. Furthermore, the mutational study of the Walker A motif, which is required for nucleotide binding and hydrolysis, showed that this motif plays indispensable roles in the function of Tih2p. When a temperature-sensitive tih2 mutant, tih2-160, was incubated at the nonpermissive temperature, cells were rapidly arrested in the G 1 phase. Northern blot analysis revealed that Tih2p is required for transcription of G 1 cyclin and of several ribosomal protein genes. The similarities between the mutant phenotypes of tih2-160 and those of taf145 mutants suggest a role for TIH2 in the regulation of RNA polymerase II-directed transcription.One of the common post-translational modifications of nuclear and cytosolic proteins in eukaryotes is the addition of N-acetylglucosamine residues O-linked (O-GlcNAc) 1 to serine and threonine residues. A number of physiologically or structurally important proteins have thus far been shown to contain O-GlcNAc, including the largest subunit of RNA polymerase II (RNAP II) (1), transcription factors such as Sp1 and hepatocyte nuclear factor 1 (2-4), nuclear pore proteins (5, 6) and chromatin-associated proteins (7). To study nuclear factors involved in nuclear transport and other nuclear events, we previously performed an in vitro binding assay of a rat liver nuclear matrix fraction using a wheat germ agglutinin affinity column (8).Several O-GlcNAc-containing proteins such as nucleoporins as well as nonglycosylated proteins like importin ␤ were isolated (9). Two RuvB-like proteins, p50 (9) and p47 (10), were also isolated by this method probably because of their interaction with O-GlcNAc-bearing proteins. RuvB is a prokaryotic DNA helicase, and its helicase activity and DNA binding affinity are enhanced by interaction with RuvA (11,12). These two factors form a large motor protein complex to promote branch migration at Holliday junctions at the late stages of homologous recombination. The p50/p47 and RuvB proteins share highly conserved Walker domains (A and B), indicative of proteins that bind nucleotide triphosphates (10, 13). Based on this, p50 and p47 were proposed to be the eukaryotic homologues of the RuvB DNA helicase. Indeed, p50 and p47 are present in a wide range of eukaryotes ranging from yeast to mammals, suggesting that this basic helicase activity may be conserved among the eukaryotes.In addition to recombination, transient unwinding of the DNA duplex is required in many cellular processes such as replication, transcription, and DNA repair. To accommodate ...

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Section: Tih2mentioning

confidence: 99%

Section: Tih2mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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The Saccharomyces cerevisiae RuvB-like Protein, Tih2p, Is Required for Cell Cycle Progression and RNA Polymerase II-directed Transcription

Lim¹,

Kimata²,

Ohdate³

et al. 2000

Journal of Biological Chemistry

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“…TIP49b (35) (synonyms: TIP48 [73], Reptin52 [3], RUVBL2 [58], Rvb2 [65)], TAP54␤ [31], and TIH2p [24]) is a protein of 463 amino acids that exhibits 43% identity with TIP49a (36) (synonyms: Pontin52 [4)], NMP238 [29)], RUVBL1 [58], Rvb1 [65)], TAP54␣ [31], and TIH1p [40]). …”

mentioning

confidence: 99%

TIP49b, a Regulator of Activating Transcription Factor 2 Response to Stress and DNA Damage

Cho

Bhoumik²,

Broday³

et al. 2001

Molecular and Cellular Biology

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Activating transcription factor 2 (ATF2/CRE-BP1) is implicated in transcriptional control150-248 in fibroblasts or melanoma but not in ATF2-null cells caused a profound G 2 M arrest and increased degree of apoptosis following irradiation. The interaction between ATF2 and TIP49b constitutes a novel mechanism that serves to limit ATF2 transcriptional activities and highlights the central role of ATF2 in the control of the cell cycle and apoptosis in response to stress and DNA damage.

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Application of immobilized ATP to the study of NLRP inflammasomes

Liao

Sandall

Carlson

et al. 2019

Archives of Biochemistry and Biophysics

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The NLRP proteins are a subfamily of the NOD-like receptor (NLR) innate immune sensors that possess an ATP-binding NACHT domain. As the most well studied member, NLRP3 can initiate the assembly process of a multiprotein complex, termed the inflammasome, upon detection of a wide range of microbial products and endogenous danger signals and results in the activation of pro-caspase-1, a cysteine protease that regulates multiple host defense pathways including cytokine maturation. Dysregulated NLRP3 activation contributes to inflammation and the pathogenesis of several chronic diseases, and the ATP-binding properties of NLRPs are thought to be critical for inflammasome activation. In light of this, we examined the utility of immobilized ATP matrices in the study of NLRP inflammasomes. Using NLRP3 as the prototypical member of the family, P-linked ATP Sepharose was determined to be a highly-effective capture agent. In subsequent examinations, P-linked ATP Sepharose was used as an enrichment tool to enable the effective profiling of NLRP3-biomarker signatures with selected reaction monitoring-mass spectrometry (SRM-MS). Finally, ATP Sepharose was used in combination with a fluorescencelinked enzyme chemoproteomic strategy (FLECS) screen to identify potential competitive inhibitors of NLRP3. The identification of a novel benzo [d]imidazol-2-one inhibitor that specifically targets the ATP-binding and hydrolysis properties of the NLRP3 protein implies that ATP Sepharose and FLECS could be applied other NLRPs as well. . The NLRP Inflammasome. The inflammasome is a protein complex and caspase-activating platform that drives inflammation and plays a crucial role in the innate immune response [1][2][3][4].Following stimulation by pathogen or danger/damage associated molecular patterns (PAMPs or DAMPs), inflammasome assembly induces the recruitment and autocatalytic cleavage of procaspases-1, 8 and 11, which are cysteine proteases and the key effectors of the inflammasome [5][6][7]. Activated caspases in turn regulate key processes involved in inflammation and host defense that includes cytokine maturation, metabolism, reactive oxygen species (ROS) generation, apoptosis and pyroptosis. For example, pro-IL-1, an endogenous pyrogen that induces fever as well as other immune responses, is cleaved by caspase-1 into mature IL-1 and is released into the extracellular environment to fulfill its physiological functions [8].Each inflammasome has scaffold proteins that determine inflammasome specificity and mediate its assembly. The nucleotide-binding oligomerization domain, leucine rich repeat and pyrin domain containing-3 (NLRP3) protein is the most extensively studied inflammasome scaffold protein. This protein belongs to the 14-member NLRP subfamily of the immune sensor Nod-like receptor (NLR) family [9]. NLRP3 possesses an N-terminal pyrin domain (PYD), a central NAIP, CIITA, HET-E and TP1 (NACHT) domain, and a C-terminal leucine rich repeat (LRR) domain [10]. The PYD of NLRP3 has been demonstrated to interact with pro-caspase-1 ...

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Molecular Shape and ATP Binding Activity of Rat p50, a Putative Mammalian Homologue of RuvB DNA Helicase

Cited by 11 publications

References 0 publications

The Saccharomyces cerevisiae RuvB-like Protein, Tih2p, Is Required for Cell Cycle Progression and RNA Polymerase II-directed Transcription

The Saccharomyces cerevisiae RuvB-like Protein, Tih2p, Is Required for Cell Cycle Progression and RNA Polymerase II-directed Transcription

TIP49b, a Regulator of Activating Transcription Factor 2 Response to Stress and DNA Damage

Application of immobilized ATP to the study of NLRP inflammasomes

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