Linker scanner mutagenesis of the Xenopus laevis ribosomal gene promoter.

R H Reeder, D Pennock, B McStay, J Roan, E Tolentino, P Walker
Author Information
  1. R H Reeder: Hutchinson Cancer Research Center, Seattle, WA 98104.

Abstract

We have assayed a series of linker scanner mutants which cover the Xenopus laevis ribosomal gene promoter at approximately ten base pair intervals. All of these mutations adversely affect promoter activity with the exception of one mutation which stimulates activity. Thus, none are neutral. We show that most of the mutations can be partially rescued by ligating a block of enhancer elements upstream of the promoter. In addition, we have made extracts from liver nuclei which produce DNaseI protection footprints over the promoter. Analysis of both strands reveals a prominent footprinting domain from about -5 to -30. However, lesser changes in the digestion pattern are detected over most of the promoter. Previously published analyses have suggested that this promoter might be composed of three functional domains. The experiments presented here suggest that either 1) the three putative domains are so closely arranged that the boundaries are difficult to discern, or 2) the situation is more complex.

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MeSH Term

Animals
Base Sequence
DNA, Recombinant
DNA, Ribosomal
Enhancer Elements, Genetic
Female
Genes, Synthetic
Molecular Sequence Data
Mutation
Oocytes
Promoter Regions, Genetic
RNA Polymerase I
RNA, Ribosomal
Xenopus laevis

Chemicals

DNA, Recombinant
DNA, Ribosomal
RNA, Ribosomal
RNA Polymerase I

Word Cloud

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