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© The Rockefeller University Press, 0021-9525/1999//1133 $5.00
The Journal of Cell Biology, Volume 145, Number 6, , 1999 1133-1143

Intron-independent Association of Splicing Factors with Active Genes

Caroline Jolly^*,, Claire Vourc'h, Michel Robert-Nicoud, and Richard I. Morimoto^*

^* Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, Evanston, Illinois 60208; and DyOGen, INSERM U309, Institut Albert Bonniot, Domaine de la Merci, 38706 La Tronche cedex, France

The cell nucleus is organized as discrete domains, often associated with specific events involved in chromosome organization, replication, and gene expression. We have examined the spatial and functional relationship between the sites of heat shock gene transcription and the speckles enriched in splicing factors in primary human fibroblasts by combining immunofluorescence and fluorescence in situ hybridization (FISH). The hsp90 and hsp70 genes are inducibly regulated by exposure to stress from a low basal level to a high rate of transcription; additionally the hsp90 gene contains 10 introns whereas the hsp70 gene is intronless. At 37°C, only 30% of hsp90 transcription sites are associated with speckles whereas little association is detected with the hsp70 gene, whose constitutive expression is undetectable relative to the hsp90 gene. Upon exposure of cells to heat shock, the heavy metal cadmium, or the amino acid analogue azetidine, transcription at the hsp90 and hsp70 gene loci is strongly induced, and both hsp transcription sites become associated with speckles in >90% of the cells. These results reveal a clear disconnection between the presence of intervening sequences at specific gene loci and the association with splicing factor–rich regions and suggest that subnuclear structures containing splicing factors are associated with sites of transcription.

Key Words: heat shock genes • nuclear organization • splicing • transcription

Abbreviations used in this paper: CTD, COOH-terminal domain; FISH, fluorescence in situ hybridization; IG, interchromatin granule; PF, perichromatin fibril; Pol II, polymerase II; snRNP, small nuclear ribonucleoprotein.

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