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Pseudouridine synthase 1: a site-specific synthase without strict sequence recognition requirements

, : Pseudouridine synthase 1: a site-specific synthase without strict sequence recognition requirements. Nucleic Acids Research 40(5): 2107-2118

Pseudouridine synthase 1 (Pus1p) is an unusual site-specific modification enzyme in that it can modify a number of positions in tRNAs and can recognize several other types of RNA. No consensus recognition sequence or structure has been identified for Pus1p. Human Pus1p was used to determine which structural or sequence elements of human tRNASer are necessary for pseudouridine () formation at position 28 in the anticodon stem-loop (ASL). Some point mutations in the ASL stem of tRNASer had significant effects on the levels of modification and compensatory mutation, to reform the base pair, restored a wild-type level of formation. Deletion analysis showed that the tRNASer TC stem-loop was a determinant for modification in the ASL. A mini-substrate composed of the ASL and TC stem-loop exhibited significant formation at position 28 and a number of mutants were tested. Substantial base pairing in the ASL stem (3 out of 5 bp) is required, but the sequence of the TC loop is not required for modification. When all nucleotides in the ASL stem other than U28 were changed in a single mutant, but base pairing was retained, a near wild-type level of modification was observed.


PMID: 22102571

DOI: 10.1093/nar/gkr1017

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