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Molecular and Cellular Biology, December 2006, p. 8710-8721, Vol. 26, No. 23
0270-7306/06/$08.00+0     doi:10.1128/MCB.01129-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

A Gene-Specific Requirement for FACT during Transcription Is Related to the Chromatin Organization of the Transcribed Region{triangledown}

Silvia Jimeno-González,1 Fernando Gómez-Herreros,1 Paula M. Alepuz,2 and Sebastián Chávez1*

Departamento de Genética, Universidad de Sevilla, E-41012 Sevilla, Spain,1 Departamento de Bioquímica y Biología Molecular, Universitat de València, E-46100 Burjassot, València, Spain2

Received 23 June 2006/ Returned for modification 27 July 2006/ Accepted 14 September 2006

The FACT complex stimulates transcription elongation on nucleosomal templates. In vivo experiments also involve FACT in the reassembly of nucleosomes traversed by RNA polymerase II. Since several features of chromatin organization vary throughout the genome, we wondered whether FACT is equally required for all genes. We show in this study that the in vivo depletion of Spt16, one of the subunits of Saccharomyces cerevisiae FACT, strongly affects transcription of three genes, GAL1, PHO5, and Kluyveromyces lactis LAC4, which exhibit positioned nucleosomes at their transcribed regions. In contrast, showing a random nucleosome structure, YAT1 and Escherichia coli lacZ are only mildly influenced by Spt16 depletion. We also show that the effect of Spt16 depletion on GAL1 expression is suppressed by a histone mutation and that the insertion of a GAL1 fragment, which allows the positioning of two nucleosomes, at the 5' end of YAT1 makes the resulting transcription unit sensitive to Spt16 depletion. These results indicate that FACT requirement for transcription depends on the chromatin organization of the 5' end of the transcribed region.

* Corresponding author. Mailing address: Departamento de Genética, Facultad de Biología, Avda. Reina Mercedes 6, 41012-Sevilla, Spain. Phone: 34-954550920. Fax: 34-954557104. E-mail: schavez{at}us.es.

{triangledown} Published ahead of print on 25 September 2006.

Molecular and Cellular Biology, December 2006, p. 8710-8721, Vol. 26, No. 23
0270-7306/06/$08.00+0     doi:10.1128/MCB.01129-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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