Article

The zinc finger protein Ynr046w is plurifunctional and a component of the eRF1 methyltransferase in yeast.

UPR 9073 du CNRS, Institut de Biologie Physico-Chimique, 13 rue Pierre et Marie Curie, 75005 Paris, France.
Journal of Biological Chemistry (impact factor: 4.77). 12/2006; 281(47):36140-8. DOI:10.1074/jbc.M608571200 pp.36140-8
Source: PubMed

ABSTRACT Protein release factor eRF1 in Saccharomyces cerevisiae, in complex with eRF3 and GTP, is methylated on a functionally crucial Gln residue by the S-adenosylmethionine-dependent methyltransferase Ydr140w. Here we show that eRF1 methylation, in addition to these previously characterized components, requires a 15-kDa zinc-binding protein, Ynr046w. Co-expression in Escherichia coli of Ynr046w and Ydr140w allows the latter to be recovered in soluble form rather than as inclusion bodies, and the two proteins co-purify on nickel-nitrilotriacetic acid chromatography when Ydr140w alone carries a His tag. The crystal structure of Ynr046w has been determined to 1.7 A resolution. It comprises a zinc-binding domain built from both the N- and C-terminal sequences and an inserted domain, absent from bacterial and archaeal orthologs of the protein, composed of three alpha-helices. The active methyltransferase is the heterodimer Ydr140w.Ynr046w, but when alone, both in solution and in crystals, Ynr046w appears to be a homodimer. The Ynr046w eRF1 methyltransferase subunit is shared by the tRNA methyltransferase Trm11p and probably by two other enzymes containing a Rossman fold.

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Keywords

15-kDa zinc-binding protein
 
active methyltransferase
 
archaeal orthologs
 
crystals
 
eRF3
 
Escherichia coli
 
functionally crucial Gln residue
 
inclusion bodies
 
methylated
 
nickel-nitrilotriacetic acid chromatography
 
Protein release factor eRF1
 
S-adenosylmethionine-dependent methyltransferase Ydr140w
 
soluble form
 
tRNA methyltransferase Trm11p
 
two proteins co-purify
 
Ynr046w eRF1 methyltransferase subunit