1 H, 13 C and 15 N resonance assignments of TFIIS LW domain from Homo sapiens
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H, 13C and 15N resonance assignments of TFIIS LW domain from Homo sapiens Han Guo1 · Jiahai Zhang1 · Jie Gao1 · Xiaoming Tu1 Received: 17 January 2020 / Accepted: 16 April 2020 © Springer Nature B.V. 2020
Abstract LW domain is the N-terminal domain I of transcription elongation factor TFIIS, which is a component of RNA polymerase II (Pol II) preinitiation complexes (PICs). Here, we report the resonance assignments of TFIIS LW domain from Homo sapiens for further understanding of the relationship between its structure and function. Keywords TFIIS · LW domain · H. sapiens · Resonance assignment
Biological context Transcription elongation factor TFIIS induces mRNA cleavage by enhancing the intrinsic nuclease activity of RNA Pol II, which passes through the template-encoded pause sites (Kettenberger et al. 2003). It is widely distributed in mammal, Drosophila, yeast and archaea (Langer and Zillig 1993). Human transcription elongation factor TFIIS (HsTFIIS) consists of three domains: a N-terminal domain I (LW domain), a central domain II, and a C-terminal domain III. HsTFIIS domain II forms a three-helix bundle and is essential for RNA Pol II to read through transcription pause sites and cleave the RNA transcripts (Morin et al. 1996). HsTFIIS domain III is nucleic acid-binding domain and the solution structure of the domain has been solved previously by Weiss and coworkers (Qian et al. 1993a, b) who refer to it as a “Zn ribbon” due to its ability to bind Zn and its β-sheet secondary structure composition. As such, domains II-III are the primary functional domains of HsTFIIS in vitro and in vivo (Kim et al. 2007). TFIIS LW domain is conserved from yeast to human and is reported to interact with the transcriptionally active Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12104-020-09945-8) contains supplementary material, which is available to authorized users. * Xiaoming Tu [email protected] 1
Hefei National Laboratory for Physical Sciences at Microscale, School of Life Sciences, University of Science and Technology of China, Anhui 230027 Hefei, China
RNA polymerase II holoenzyme (Pan et al. 1997; Booth et al. 2000). To gain insight into the structure and other functions of HsTFIIS LW domain, we analyzed the NMR data of HsTFIIS LW domain to establish its structural information. Here, we report the backbone and side-chain 1H, 13C and 15N resonance assignments of HsTFIIS LW domain. Residues of this domain were almost completely identified: a total of 98% of backbone and 98% side-chain assignments have been obtained.
Methods and experiments The DNA fragment encoding the LW domain (residue 1–96) of HsTFIIS (UniProt accession number P23193) was obtained by PCR from human genomic DNA using the two primers 5′-CATACATATGGAGGACGAAGTGGTCCGC3′ and 5′-CACGCTCGAGTCATGTAATTGCAGGTTCTT TC-3′ and cloned into the NdeI/XhoI-cleaved vector pET28a (Novagen). The recombinant vector was then transformed and overproduced into Escherichia coli strain BL21
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