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Related Experiment Videos

RNA-dependent cysteine biosynthesis in archaea.

Anselm Sauerwald1, Wenhong Zhu, Tiffany A Major

  • 1Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520-8114, USA.

Science (New York, N.Y.)
|March 26, 2005
PubMed
Summary

Methanogenic archaea use a novel pathway for cysteine synthesis, employing O-phosphoseryl-tRNA synthetase (SepRS) and Sep-tRNA:Cys-tRNA synthase (SepCysS) to create Cys-tRNA(Cys). This pathway is crucial for cysteine biosynthesis in organisms lacking cysteinyl-tRNA synthetase (CysRS).

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Archaea Biology

Background:

  • Many methanogenic archaea lack the canonical cysteinyl-tRNA synthetase (CysRS).
  • This suggests an alternative mechanism for cysteine incorporation into proteins exists in these organisms.

Purpose of the Study:

  • To elucidate the mechanism of cysteine-tRNA(Cys) formation in archaea lacking CysRS.
  • To investigate the role of O-phosphoserine (Sep) in this alternative pathway.
  • To determine if this pathway is essential for cysteine biosynthesis.

Main Methods:

  • Enzyme activity assays using partially purified proteins from Methanocaldococcus jannaschii.
  • Identification and characterization of O-phosphoseryl-tRNA synthetase (SepRS) and Sep-tRNA:Cys-tRNA synthase (SepCysS).

Related Experiment Videos

  • Comparative genomic analysis across archaeal species.
  • Gene deletion experiments in Methanococcus maripaludis to assess cysteine auxotrophy.

    • Main Results:

    • tRNA(Cys) is acylated with O-phosphoserine (Sep) by SepRS, not cysteine.
    • Sep-tRNA(Cys) is subsequently converted to Cys-tRNA(Cys) by SepCysS.
    • The SepRS-SepCysS pathway is essential for cysteine biosynthesis in Methanococcus maripaludis, as demonstrated by auxotrophy upon gene deletion.
    • Genomic data indicates this pathway is conserved in all organisms lacking CysRS.

    Conclusions:

    • A novel pathway involving SepRS and SepCysS is responsible for Cys-tRNA(Cys) synthesis in CysRS-deficient archaea.
    • This pathway is essential for cysteine biosynthesis and protein translation in these organisms.
    • The chemical analogy between Sep-tRNA conversions and selenocysteine incorporation suggests this pathway may be ancestral for both amino acids in the genetic code.