tRNASer(CGA) differentially regulates expression of wild-type and codon-modified papillomavirus L1 genes.
Exogenous transfer RNAs (tRNAs) favor translation of bovine papillomavirus 1 wild-type (wt) L1 mRNA in in vitro translation systems (Zhou et al. 1999, J. Virol., 73, 4972-4982). We, therefore, investigated whether papillomavirus (PV) wt L1 protein expression could be enhanced in eukaryotic cells following exogenous tRNA supplementation. Both Chinese hamster ovary (CHO) and Cos1 cells, transfected with PV1 wt L1 genes, effectively transcribed the genes but did not translate them. However, L1 protein translation was demonstrated following co-transfection with the L1 gene and a gene expressing tRNA(Ser)(CGA). Cell lines, stably transfected with a bovine papillomavirus 1 (BPV1) wt L1 expression construct, produced L1 protein after the transfection of the tRNA(Ser)(CGA) gene, but not following the transfection with basal vectors, suggesting that tRNA(Ser)(CGA) gene enhanced wt L1 translation as a result of endogenous tRNA alterations and phosphorylation of translation initiation factors elF4E and elF2alpha in the tRNA(Ser)(CGA) transfected L1 cell lines. The tRNA(Ser)(CGA) gene expression significantly reduced translation of L1 proteins expressed from codon-modified (HB) PV L1 genes utilizing mammalian preferred codons, but had variable effects on translation of green fluorescent proteins (GFPs) expressed from six serine GFP variants. The changes of tRNA pools appear to match the codon composition of PV wt and HB L1 genes and serine GFP variants to regulate translation of their mRNAs. These findings demonstrate for the first time in eukaryotic cells that translation of the target genes can be differentially influenced by the provision of a single tRNA expression construct.
Gu
Wenyi
W
Centre for Immunology and Cancer Research, University of Queensland, Princess Alexandra Hospital, Woolloongabba, Queensland 4102, Australia.
Li
Mengrong
M
Zhao
Wei Ming
WM
Fang
Ning Xia
NX
Bu
Shurui
S
Frazer
Ian H
IH
Zhao
Kong-Nan
KN
D000818:Animals; D052688:Bovine papillomavirus 1 / Q000235:genetics; D016466:CHO Cells; D019556:COS Cells; D036022:Capsid Proteins / Q000235:genetics* / Q000378:metabolism; D002522:Chlorocebus aethiops; D003062:Codon; D006224:Cricetinae; D003412:Cricetulus; D015967:Gene Expression Regulation, Viral; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D010448:Peptide Initiation Factors / Q000378:metabolism; D010766:Phosphorylation; D014176:Protein Biosynthesis; D012343:RNA, Transfer / Q000378:metabolism; D012362:RNA, Transfer, Ser / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
D036022:Capsid Proteins; D003062:Codon; C085217:L1 protein, Bovine papillomavirus; D008164:Luminescent Proteins; D010448:Peptide Initiation Factors; D012362:RNA, Transfer, Ser; D049452:Green Fluorescent Proteins; D012343:RNA, Transfer