title: In vitro processing of the human growth hormone primary transcript.
abstract: To study the sequence of events during processing of primary RNA transcripts and to gain more insight into the mechanism of splice site selection, the in vitro processing of a 2.5 kb human growth hormone (hGH) pre-mRNA containing four introns and an alternative 3' splice site for intron B was analysed. In order to process the hGH pre-mRNA the preparation of the HeLa cell nuclear extract had to be modified, indicating differences in factor requirement for processing this pre-mRNA. After an unusual long lag phase of one hour splicing intermediates begin to accumulate. Intron A and D are removed with correct ligation of exons 1/2 and 4/5. Most splice sites are used--albeit with variable efficiencies--except the splice sites surrounding exon 3 and the 3' alternative splice site within exon 3; as a consequence "exon skipping" events take place. Using a pre-mRNA containing only intron B neither the 5' nor the 3' splice site is cleaved, indicating that the 3' splice site of intron B is not recognized. The results show that splice sites can differ considerably in their requirement for splicing factors.
authors: Institut für Biochemie, Universität Wien, Austria.
authors: K
authors: K
authors: Hartmuth
authors: A
authors: A
authors: Barta
mesh_terms: D003001:Cloning, Molecular; D005091:Exons; D013006:Growth Hormone / Q000235:genetics*; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D007438:Introns; D009710:Nucleotide Mapping; D010957:Plasmids; D012322:RNA Precursors / Q000235:genetics; D012323:RNA Processing, Post-Transcriptional; D012326:RNA Splicing; D014158:Transcription, Genetic
chemical_list: D012322:RNA Precursors; D013006:Growth Hormone