8SZS

Cat DHX9 bound to GDP

Summary for 8SZS

• Entry DOI: 10.2210/pdb8szs/pdb
• Descriptor: RNA helicase, GUANOSINE-5'-DIPHOSPHATE, MAGNESIUM ION, ... (6 entities in total)
• Functional Keywords: dexh-box, rha, hydrolase
• Biological source: Felis catus (domestic cat)
• Total number of polymer chains: 1
• Total formula weight: 116992.67

Authors

Lee, Y.-T.,Sickmier, E.A.,Boriack-Sjodin, P.A. (deposition date: 2023-05-30, release date: 2023-09-06, Last modification date: 2023-12-27)

Primary citation

Gotur, D.,Case, A.,Liu, J.,Sickmier, E.A.,Holt, N.,Knockenhauer, K.E.,Yao, S.,Lee, Y.T.,Copeland, R.A.,Buker, S.M.,Boriack-Sjodin, P.A.
Development of assays to support identification and characterization of modulators of DExH-box helicase DHX9.
Slas Discov, 28:376-384, 2023

PubMed Abstract: DHX9 is a DExH-box RNA helicase that utilizes hydrolysis of all four nucleotide triphosphates (NTPs) to power cycles of 3' to 5' directional movement to resolve and/or unwind double stranded RNA, DNA, and RNA/DNA hybrids, R-loops, triplex-DNA and G-quadraplexes. DHX9 activity is important for both viral amplification and maintaining genomic stability in cancer cells; therefore, it is a therapeutic target of interest for drug discovery efforts. Biochemical assays measuring ATP hydrolysis and oligonucleotide unwinding for DHX9 have been developed and characterized, and these assays can support high-throughput compound screening efforts under balanced conditions. Assay development efforts revealed DHX9 can use double stranded RNA with 18-mer poly(U) 3' overhangs and as well as significantly shorter overhangs at the 5' or 3' end as substrates. The enzymatic assays are augmented by a robust SPR assay for compound validation. A mechanism-derived inhibitor, GTPγS, was characterized as part of the validation of these assays and a crystal structure of GDP bound to cat DHX9 has been solved. In addition to enabling drug discovery efforts for DHX9, these assays may be extrapolated to other RNA helicases providing a valuable toolkit for this important target class.

PubMed: 37625785
DOI: 10.1016/j.slasd.2023.08.006

Experimental method

X-RAY DIFFRACTION (2.38 Å)