Improved crystals of<i>Thermus thermophilus</i>prolyl-tRNA synthetase complexed with cognate tRNA obtained by crystallization from precipitate

Yaremchuk, A.; Kriklivyi, I. A.; Cusack, Stephen; Tukalo, M. A.

doi:10.1107/s0907444999015504

Cited by 10 publications

(24 citation statements)

References 13 publications

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“…The structure of a ProRSTT–tRNA Pro complex has been previously described at 3.5 Å resolution (Cusack et al ., 1998). A closely related crystal form of the same complex was later grown with superior diffraction quality, permitting data to be collected up to 2.85 Å resolution (Yaremchuk et al ., 2000b). Here we describe in more detail the structure of the ProRSTT–tRNA Pro complex.…”

Section: Resultsmentioning

confidence: 99%

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Crystal structure of a eukaryote/archaeon-like prolyl-tRNA synthetase and its complex with tRNAPro(CGG)

Yaremchuk

2000

The EMBO Journal

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Prolyl‐tRNA synthetase (ProRS) is a class IIa synthetase that, according to sequence analysis, occurs in different organisms with one of two quite distinct structural architectures: prokaryote‐like and eukaryote/archaeon‐like. The primary sequence of ProRS from the hypothermophilic eubacterium Thermus thermophilus (ProRSTT) shows that this enzyme is surprisingly eukaryote/archaeon‐like. We describe its crystal structure at 2.43 Å resolution, which reveals a feature that is unique among class II synthetases. This is an additional zinc‐containing domain after the expected class IIa anticodon‐binding domain and whose C‐terminal extremity, which ends in an absolutely conserved tyrosine, folds back into the active site. We also present an improved structure of ProRSTT complexed with tRNAPro(CGG) at 2.85 Å resolution. This structure represents an initial docking state of the tRNA in which the anticodon stem–loop is engaged, particularly via the tRNAPro‐specific bases G35 and G36, but the 3′ end does not enter the active site. Considerable structural changes in tRNA and/or synthetase, which are probably induced by small substrates, are required to achieve the conformation active for aminoacylation.

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Section: Resultsmentioning

confidence: 99%

“…Crystallization of the ProRSTT–tRNA Pro complex is described elsewhere (Yaremchuk et al ., 2000b). Crystal were cryoprotected with 27% glycerol.…”

Section: Methodsmentioning

confidence: 99%

Crystal structure of a eukaryote/archaeon-like prolyl-tRNA synthetase and its complex with tRNAPro(CGG)

Yaremchuk

2000

The EMBO Journal

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“…In the absence of antibodies against the two enzymes, we attempted to inhibit cysteinylation with the proline analogs, which are not expected to inhibit the canonical CysRS enzyme that may also be expressed in G. lamblia. As can be seen, under the conditions tested, the major CysRS activity derives from ProCysRS, as it can be inhibited by thiaproline (24) and (25). However, the inhibition is not as good as with the pure ProCysRS; thus, there may be a small amount of canonical CysRS activity in G. lamblia.…”

Section: G Lamblia Prors Synthesizes Cys-trna In Vivomentioning

confidence: 90%

A dual-specificity aminoacyl-tRNA synthetase in the deep-rooted eukaryote Giardia lamblia

Bunjun

Stathopoulos

Graham

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

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Cysteinyl-tRNA (Cys-tRNA) is essential for protein synthesis. In most organisms the enzyme responsible for the formation of Cys-tRNA is cysteinyl-tRNA synthetase (CysRS). The only known exceptions are the euryarchaea Methanococcus jannaschii and Methanobacterium thermoautotrophicum, which do not encode a CysRS. Deviating from the accepted concept of one aminoacyltRNA synthetase per amino acid, these organisms employ prolyltRNA synthetase as the enzyme that carries out Cys-tRNA formation. To date this dual-specificity prolyl-cysteinyl-tRNA synthetase (ProCysRS) is only known to exist in archaea. Analysis of the preliminary genomic sequence of the primitive eukaryote Giardia lamblia indicated the presence of an archaeal prolyl-tRNA synthetase (ProRS). Its proS gene was cloned and the gene product overexpressed in Escherichia coli. By using G. lamblia, M. jannaschii, or E. coli tRNA as substrate, this ProRS was able to form Cys-tRNA and Pro-tRNA in vitro. Cys-AMP formation, but not Pro-AMP synthesis, was tRNA-dependent. The in vitro data were confirmed in vivo, as the cloned G. lamblia proS gene was able to complement a temperature-sensitive E. coli cysS strain. Inhibition studies of CysRS activity with proline analogs (thiaproline and 5-O-[N-(Lprolyl)-sulfamoyl]adenosine) in a Giardia S-100 extract predicted that the organism also contains a canonical CysRS. This prediction was confirmed by cloning and analysis of the corresponding cysS gene. Like a number of archaea, Giardia contains two enzymes, ProCysRS and CysRS, for Cys-tRNA formation. In contrast, the purified Saccharomyces cerevisiae and E. coli ProRS enzymes were unable to form Cys-tRNA under these conditions. Thus, the dual specificity is restricted to the archaeal genre of ProRS. G. lamblia's archaeal-type prolyl-and alanyl-tRNA synthetases refine our understanding of the evolution and interaction of archaeal and eukaryal translation systems.

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“…For example, a prolyl-tRNA synthetase (PARS) homodimer of Thermus thermophilus with the cognate tRNA, as well as the human PARS homodimer with halofuginone (a dual-site inhibitor for tRNA and amino acid binding) and ATP, are captured with the asymmetric unit. 10,11 Likewise, yeast tRNA Asp functionally interconnects the active-site domain of one monomer and the anticodon-binding region of the other monomer of Escherichia coli aspartyl-tRNA synthetase (DARS) homodimer. 12 Furthermore, in humans, C-terminus of QARS1, N-terminus of RARS1, and N-terminus of AIMP1 form a tertiary subcomplex bearing the asymmetric unit, which is able to undergo rigid-body rotational motion to facilitate binding of tRNA.…”

Section: Introductionmentioning

confidence: 99%

Cell‐based analysis of pairwise interactions between the components of the multi‐tRNA synthetase complex

Kong

Kim

2020

The FASEB Journal

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Improved crystals ofThermus thermophilusprolyl-tRNA synthetase complexed with cognate tRNA obtained by crystallization from precipitate

Cited by 10 publications

References 13 publications

Crystal structure of a eukaryote/archaeon-like prolyl-tRNA synthetase and its complex with tRNAPro(CGG)

Crystal structure of a eukaryote/archaeon-like prolyl-tRNA synthetase and its complex with tRNAPro(CGG)

A dual-specificity aminoacyl-tRNA synthetase in the deep-rooted eukaryote Giardia lamblia

Cell‐based analysis of pairwise interactions between the components of the multi‐tRNA synthetase complex

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