title: Disruption and complementation of the selenocysteine biosynthesis pathway reveals a hierarchy of selenoprotein gene expression in the archaeon Methanococcus maripaludis.
abstract: Proteins containing selenocysteine are found in members of all three domains of life, Bacteria, Eukarya and Archaea. A dedicated tRNA (tRNA(sec)) serves as a scaffold for selenocysteine synthesis. However, sequence and secondary structures differ in tRNA(sec) from the different domains. An Escherichia coli strain lacking the gene for tRNA(sec) could only be complemented with the homologue from Methanococcus maripaludis when a single base in the anticodon loop was exchanged demonstrating that this base is a crucial determinant for archaeal tRNA(sec) to function in E. coli. Complementation in trans of M. maripaludis JJ mutants lacking tRNA(sec) , O-phosphoseryl-tRNA(sec) kinase or O-phosphoseryl-tRNA(sec) :selenocysteine synthase with the corresponding genes from M. maripaludis S2 restored the mutant's ability to synthesize selenoproteins. However, only partial restoration of the wild-type selenoproteome was observed as only selenocysteine-containing formate dehydrogenase was synthesized. Quantification of transcripts showed that disrupting the pathway of selenocysteine synthesis leads to downregulation of selenoprotein gene expression, concomitant with upregulation of a selenium-independent backup system, which is not re-adjusted upon complementation. This transcriptional arrest was independent of selenophosphate but depended on the 'history' of the mutants and was inheritable, which suggests that a stable genetic switch may cause the resulting hierarchy of selenoproteins synthesized.
authors: Institut für Molekulare Biowissenschaften, Johann Wolfgang Goethe-Universität, Frankfurt am Main, Germany.
authors: Tilmann
authors: T
authors: Stock
authors: Mirjam
authors: M
authors: Selzer
authors: Sarah
authors: S
authors: Connery
authors: Deniz
authors: D
authors: Seyhan
authors: Armin
authors: A
authors: Resch
authors: Michael
authors: M
authors: Rother
mesh_terms: D053898:Biosynthetic Pathways / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D017353:Gene Deletion; D020869:Gene Expression Profiling; D019848:Gene Expression Regulation, Archaeal; D005816:Genetic Complementation Test; D017017:Methanococcus / Q000235:genetics* / Q000378:metabolism; D017279:Selenocysteine / Q000096:biosynthesis*; D051140:Selenoproteins / Q000096:biosynthesis*; D014158:Transcription, Genetic
chemical_list: D051140:Selenoproteins; D017279:Selenocysteine