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Nucleic Acids Research, 1984, Vol. 12, No. 10 4397-4410
© 1984

MOLECULAR BIOLOGY

Characterization of the size and 5' cap of messenger RNA encoding neurophysin precursors

Christopher J. Hough and Irwin M. Chaiken

Laboratory of Cbemical Biology, National Institute of Arthritis, Diabetes, and Digestive and Kidney Diseases, National Institutes of Health Bethesda, MD 20205, USA

Received December 12, 1983. Revised April 12, 1984. Accepted April 12, 1984.

The coding activity of bovine hypothalamic poly A+ mRNA for neurophysin I and II immunoreactive proteins was charscterized with respect to size and 5' cap. The mRNA was fractionated by methylmercuric hydroxide agarose gel electrophoresis and subsequently translated in vitro in rabbit reticulocyte lysates. Alternatively, nRNA was fractionated by gel exclusion HPLC and translated in wheat germ extracts. Imnunoprecipitated translation products were analyzed by gel exclusion HPLC. Neurophysin-immunoapecific protein of ~17,000 daltons, the size expected for the neuropeptide hormone-neurophysin precursors, was encoded by mRNA species of two size classes. The smaller class of mRNA's was of the size expected from the size of the precursor proteins. The larger class was 5–10 times larger. The low K+ concentration optimum for translation of unfractionated mRNA encoding neurophysin I-immunoreactive proteins and the inability of a cap analogue to inhibit this translation suggest that mRNA species encoding neurophysin I-immunoreactive translation products are incompletely capped. By contrast, the mRNA encoding neurophysin II-immunoreactive products appear to contain a normal cap structure.


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