A Yeast Chromosomal Origin of DNA Replication Defined by Multiple Functional Elements

Science. 1992 Feb 14;255(5046):817-23. doi: 10.1126/science.1536007.

Abstract

Although it has been demonstrated that discrete origins of DNA replication exist in eukaryotic cellular chromosomes, the detailed organization of a eukaryotic cellular origin remains to be determined. Linker substitution mutations were constructed across the entire Saccharomyces cerevisiae chromosomal origin, ARS1. Functional studies of these mutants revealed one essential element (A), which includes a match to the ARS consensus sequence, and three additional elements (B1, B2, and B3), which collectively are also essential for origin function. These four elements arranged exactly as in ARS1, but surrounded by completely unrelated sequence, functioned as an efficient origin. Element B3 is the binding site for the transcription factor-origin binding protein ABF1. Other transcription factor binding sites substitute for the B3 element and a trans-acting transcriptional activation domain is required. The multipartite nature of a chromosomal replication origin and the role of transcriptional activators in its function present a striking similarity to the organization of eukaryotic promoters.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Chromosome Mapping
  • Chromosomes, Fungal / chemistry
  • DNA Replication*
  • DNA, Fungal / physiology*
  • DNA-Binding Proteins*
  • Fungal Proteins / genetics
  • Gene Expression Regulation, Fungal
  • Genetic Linkage
  • Molecular Sequence Data
  • Mutation
  • Plasmids
  • Replicon / physiology
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins*
  • Trans-Activators / physiology
  • Transcription Factors*
  • Transformation, Genetic

Substances

  • ABF1 protein, S cerevisiae
  • DNA, Fungal
  • DNA-Binding Proteins
  • Fungal Proteins
  • Saccharomyces cerevisiae Proteins
  • Trans-Activators
  • Transcription Factors