Genetics, Vol. 162, 615-632, October 2002, Copyright © 2002

The Ubiquitin-Dependent Targeting Pathway in Saccharomyces cerevisiae Plays a Critical Role in Multiple Chromatin Assembly Regulatory Steps

Troy A. A. Harknessa,b, Gerald F. Daviesa, Vijay Ramaswamyb, and Terra G. Arnasonb
a Department of Anatomy and Cell Biology, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5E5, Canada
b Department of Biochemistry, University of Alberta, Edmonton, Alberta T6G 2H7, Canada

Corresponding author: Troy A. A. Harkness, College of Medicine, University of Saskatchewan, B313 Health Sciences Bldg., 107 Wiggins Rd., Saskatoon, Saskatchewan S7N 5E5, Canada., troy.harkness{at}usask.ca (E-mail)

Communicating editor: F. WINSTON

In a screen designed to isolate Saccharomyces cerevisiae strains defective for in vitro chromatin assembly, two temperature-sensitive (ts) mutants were obtained: rmc1 and rmc3 (remodeling of chromatin). Cloning of RMC1 and RMC3 revealed a broad role for the ubiquitin-dependent targeting cascade as the ubiquitin-protein ligases (E3s), the anaphase promoting complex (APC; RMC1 encodes APC5) and Rsp5p, respectively, were identified. Genetic studies linked the rmc1/apc5 chromatin assembly defect to APC function: rmc1/apc5 genetically interacted with apc9{Delta}, apc10{Delta}, and cdc26{Delta} mutants. Furthermore, phenotypes associated with the rmc1/apc5 allele were consistent with defects in chromatin metabolism and in APC function: (i) UV sensitivity, (ii) plasmid loss, (iii) accumulation of G2/M cells, and (iv) suppression of the ts defect by growth on glucose-free media and by expression of ubiquitin. On the other hand, the multifunctional E3, Rsp5p, was shown to be required for both in vitro and in vivo chromatin assembly, as well as for the proper transcriptional and translational control of at least histone H3. The finding that the distinctly different E3 enzymes, APC and Rsp5p, both play roles in regulating chromatin assembly highlight the depth of the regulatory networks at play. The significance of these findings will be discussed.




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