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Molecular and Cellular Biology, January 2004, p. 398-406, Vol. 24, No. 1
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.1.398-406.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Transcription Termination Factor reb1p Causes Two Replication Fork Barriers at Its Cognate Sites in Fission Yeast Ribosomal DNA In Vivo

Alicia Sánchez-Gorostiaga, Carlos López-Estraño,{dagger} Dora B. Krimer, Jorge B. Schvartzman, and Pablo Hernández*

Departamento de Biología Celular y del Desarrollo, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, 28040 Madrid, Spain

Received 7 July 2003/ Returned for modification 11 August 2003/ Accepted 25 September 2003

Polar replication fork barriers (RFBs) near the 3' end of the rRNA transcriptional unit are a conserved feature of ribosomal DNA (rDNA) replication in eukaryotes. In the mouse, in vivo studies indicate that the cis-acting Sal boxes required for rRNA transcription termination are also involved in replication fork blockage. On the contrary, in the budding yeast Saccharomyces cerevisiae, the rRNA transcription termination factors are not required for RFBs. Here we characterized the rDNA RFBs in the fission yeast Schizosaccharomyces pombe. S. pombe rDNA contains three closely spaced polar replication barriers named RFB1, RFB2, and RFB3 in the 3' to 5' order. The transcription termination protein reb1 and its two binding sites, present at the 3' end of the coding region, were required for fork arrest at RFB2 and RFB3 in vivo. On the other hand, fork arrest at the strongest RFB1 barrier was independent of the above transcription termination factors. Therefore, RFB2 and RFB3 resemble the barriers present in the mouse rDNA, whereas RFB1 is similar to the budding yeast RFBs. These results suggest that during evolution, cis- and trans-acting factors required for rRNA transcription termination became involved in replication fork blockage also. S. pombe is suggested to be a transitional species in which both mechanisms coexist.

* Corresponding author. Mailing address: Departamento de Biología Celular y del Desarrollo, Centro de Investigaciones Biológicas (CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain. Phone: 34-918373112, ext. 4240. Fax: 34-5360432. E-mail: p.hernandez{at}cib.csic.es.

{dagger} Present address: Department of Pathology, Medical School, Northwestern University, Chicago, IL 60610.

Molecular and Cellular Biology, January 2004, p. 398-406, Vol. 24, No. 1
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.1.398-406.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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