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Nucleic Acids Research, 1977, Vol. 4, No. 6 2009-2020
© 1977

Articles

Rabbit liver tRNA1Val: II. Unusual secondary structure of T{Psi}C stem and loop due to a U54:A60 base pair

Peter Jank, Detlev Riesner+ and Hans J. Gross

Max-Planck-lnst. fuür Biochem. D-8033 Martinsried bei Muünchen +Inst. fuür Klin. Biochem. und Physiol. Chem., Med. Hochschule Hannover D-3000 Hannover-Kleefeld, GFR

Received March 29, 1977. In contrast to all other known tRNAs, mammalian Formula contains two adenosines A59 and A60, opposite to U54 and {Psi}55 in the U{Psi}CG sequence of the T{Psi}C loop, which could form unusual A:U (or A:{Psi}) pairs in addition to the five "normal" G:C pairs. In order to measure the number of G:C and A:U (A:{Psi} ) pairs in the T{Psi}C stem, we prepared the 30 nucleotide long 3-terminal fragment of this tRNA by "m7G-cleavage". From differentiated melting curves and temperature jump experiments it was concluded that the T{Psi}C stem in this fragment is in fact extended by an additional A60:U54 pair. A dimer of this fragment with 14 base pairs was characterized by gel electrophoresis and by the same physical methods. An additional A:U pair in the Formula fragment does not necessarily mean that this is also true for intact tRNA. However, we showed that U54 is far less available for enzymatic methylatlon in mammalian Formula compared to tRNA from T- E. coli. This clear difference in U54 reactivity, together with the identification of an extra A60:U54 pair in the U{Psi}CG containing fragment suggests the presence of a 6 base pair T{Psi}C stem and a 5 nucleotide T{Psi}C loop in this tRNA.


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