2014
DOI: 10.1093/nar/gku087
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Structure of a rare non-standard sequence k-turn bound by L7Ae protein

Abstract: Kt-23 from Thelohania solenopsae is a rare RNA kink turn (k-turn) where an adenine replaces the normal guanine at the 2n position. L7Ae is a member of a strongly conserved family of proteins that bind a range of k-turn structures in the ribosome, box C/D and H/ACA small nucleolar RNAs and U4 small nuclear RNA. We have solved the crystal structure of T. solenopsae Kt-23 RNA bound to Archeoglobus fulgidus L7Ae protein at a resolution of 2.95 Å. The protein binds in the major groove displayed on the outer face of… Show more

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Cited by 17 publications
(16 citation statements)
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“…A. fulgidus L7Ae protein was expressed and purified as previously described 50 . Briefly, a modified pET-Duet1 plasmid (Novagen) containing gene sequence of a hexahistidine-L7Ae fusion protein was expressed in E. coli BL21-Gold (DE3) pLysS cells (Stratagene), induced with 0.2 mM IPTG at 20 °C for 12 h. Harvested cells were suspended in 20 mM Tris-HCl, (pH 8.0), 500 mM NaCl, 10 mM imidazole, 1 mM phenylmethylsulfonyl fluoride (PMSF) (buffer A) and lysed by sonication.…”
Section: Methodsmentioning
confidence: 99%
“…A. fulgidus L7Ae protein was expressed and purified as previously described 50 . Briefly, a modified pET-Duet1 plasmid (Novagen) containing gene sequence of a hexahistidine-L7Ae fusion protein was expressed in E. coli BL21-Gold (DE3) pLysS cells (Stratagene), induced with 0.2 mM IPTG at 20 °C for 12 h. Harvested cells were suspended in 20 mM Tris-HCl, (pH 8.0), 500 mM NaCl, 10 mM imidazole, 1 mM phenylmethylsulfonyl fluoride (PMSF) (buffer A) and lysed by sonication.…”
Section: Methodsmentioning
confidence: 99%
“…This behavior is not unique; we have found equivalent behavior for a K-turn with an A·A pair at the 2b·2n position. We have recently solved a crystal structure of the T. solenopsae Kt-23 as a duplex bound by L7Ae protein, where it adopts an N3 structure [40] . This clearly contrasts with the N1 class structure for the exact same sequence located in the SAM-I riboswitch [26] .…”
Section: The N3 and N1 Class K-turnsmentioning
confidence: 99%
“…Such adjustment seems possible, and it is observed in some homologs with known structures. A good example is U1A RRM crystallized in complex with RNA (PDB code 4C4W (Huang & Lilley, 2014)). In this structure, the RRM domains that do not bind RNA have their C-terminal α-helix perfectly overlapping with the C-terminal α-helix of AcRRM (Fig.…”
Section: Manuscript To Be Reviewed Discussionmentioning
confidence: 99%
“…Superposition of AcRRM with the RRM of U1A protein (PDB code 4c4w(Huang & Lilley, 2014)):(A) without RNA (B) in complex with RNA (dark brown color), with demonstration of the Cterminal α-helix relocation upon RNA binding. AcRRM is colored as inFigure 2, and U1A in light grey with C-terminal α-helix highlighted in dark grey.…”
mentioning
confidence: 99%