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Nucleic Acids Research, 1990, Vol. 18, No. 4 703-710
© 1990

MOLECULAR BIOLOGY

Heterogeneities in vertebrate tRNAsTrp Avian retroviruses package only as a primer the tRNATrp lacking modified m2G in positon 7

Gérard Keith and Thérèse Heyman1

Institut de Biologie Moléculaire et Cellulaire du CNRS and Centre de Recherches de Biologie Moléculaire et Cellulaire de I'Université Louis Pasteur 15 rue René Descartes, F67084 Strasbourg 1Institut Cune, Section de Biologie, Centre Universitaire F91405 Orsay, France

Received December 19, 1989. Revised January 23, 1990. Accepted January 23, 1990.

Bovine, rabbit and chicken tRNATrp species and tRNATrP packaged in avian myeloblastosis virus were separated and purified using twodimensionai gel electrophoresis and their primary structures were determined. Two major tRNATrp species (1 and 2) were identified in beef and rabbit, two minor ones (3 and 4) in beef and only one minor in rabbit. Their structures differ by 4 nucleotide substitutions located in the D, and T loops (positions 16, 47, 57 and 59). Species and 4 differ from one another by only one nucleotide at position 2. Differences between tRNATrp species were also observed in the extent of methylation of some nucleotides. Chicken tRNATrp presents only one species similar to the mammalian typel tRNATrp. In the case of the three studied animals this tRNA could be separated Into two subspecies, which differ by a post- transcriptional modification of nucleotide 7 in the acceptor stem : G or m2G. However only the non methylated species is used as the primer of DNA-RNA directed retrotranscription since it is only that form which was found in avian retroviruses. The methylation of G to m2G at position 7 could thus prevent the recognition of tRNATFP by retrovirai protein(s) responsible for the selective packaging of the primer tRNATrp.


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