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Nucleic Acids Research, 2002, Vol. 30, No. 21 4592-4598
© 2002 Oxford University Press

Temperature-dependent splicing of ß-globin pre-mRNA

Federica Gemignani, Peter Sazani, Paul Morcos1 and Ryszard Kole*

Lineberger Comprehensive Cancer Center and Department of Pharmacology, CB 7295, University of North Carolina, Chapel Hill, NC 27599-7295, USA and 1 GeneTools Inc., Philomath, OR 97370, USA

*To whom correspondence should be addressed. Tel: +1 919 966 1143; Fax: +1 919 966 3015; Email: kole{at}med.unc.edu

A T->G mutation at nucleotide 705 of human ß-globin intron 2 creates an aberrant 5' splice site and activates a cryptic 3' splice site upstream. In consequence, the pre-mRNA is spliced via aberrant splice sites, despite the presence of the still functional correct sites. Surprisingly, when IVS2-705 HeLa or K562 cells were cultured at temperatures below 30°C, aberrant splicing was inhibited and correct splicing was restored. Similar temperature effects were seen for another ß-globin pre-mRNA, IVS2-745, and in a construct in which a ß-globin intron was inserted into a coding sequence of EGFP. Temperature-induced alternative splicing was affected by the nature of the internal aberrant splice sites flanking the correct sites and by exonic sequences. The results indicate that in the context of thalassemic splicing mutations and possibly in other alternatively spliced pre-mRNAs, temperature is one of the parameters that affect splice site selection.


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