Nucleic Acids Research, 2000, Vol. 28, No. 12 2375-2382
© 2000 Oxford University Press
A gender-specific mRNA encoding a cytotoxic ribonuclease contains a 3' UTR of unusual length and structure
Laboratory of Biochemical Physiology and 1Laboratory of Experimental and Computational Biology, Division of Basic Science and 2Intramural Research Support Program, SAIC Frederick, National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, MD 21702, USA
A cDNA (2855 nt) encoding a putative cytotoxic ribonuclease (rapLR1) related to the antitumor protein onconase was cloned from a library derived from the liver of gravid female amphibian Rana pipiens. The cDNA was mainly comprised (83%) of 3' untranslated region (UTR). Secondary structure analysis predicted two unusual folding regions (UFRs) in the RNA 3' UTR. Two of these regions (711–1442 and 1877–2130 nt) contained remarkable, stalk-like, stem–loop structures greater than 38 and 12 standard deviations more stable than by chance, respectively. Secondary structure modeling demonstrated similar structures in the 3' UTRs of other species at low frequencies (0.01–0.3%). The size of the rapLR1 cDNA corresponded to the major hybridizing RNA cross-reactive with a genomic clone encoding onconase (3.6 kb). The transcript was found only in liver mRNA from female frogs. In contrast, immunoreactive onconase protein was detected only in oocytes. Deletion of the 3' UTR facilitated the in vitro translation of the rapLR1 cDNA. Taken together these results suggest that these unusual UFRs may affect mRNA metabolism and/or translation.
* To whom correspondence should be addressed. Tel: +1 301 846 5532; Fax: +1 301 846 5598; Email: jmaizel@ncifcrf.gov Present address: Susanna M. Rybak, Developmental Therapeutics Program, Division of Cancer Treatment and Diagnosis, National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, MD 21702, USA The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors
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