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Nucleic Acids Research, 1990, Vol. 18, No. 20 6031-6037
© 1990

MOLECULAR BIOLOGY

Chromosomal footprinting of transcriptionally active and inactive oocyte-type 5S RNA genes of Xenopus laevis

David R. Engelke* and Joel M. Gottesfeld1

Department of Biological Chemistry, University of Michigan Ann Arbor, Ml 48109, USA 1Division of Developmental Biology, Medical Biology Institute La Jolla, CA 92037, USA

*To whom correspondence should be addressed

Received June 28, 1990. Revised September 5, 1990. Accepted September 5, 1990.

The chromatin structure of the Xenopus oocyte-speciflc 5S rRNA genes was examined at high resolution in immature oocyte and somatic cell chromosomes by DNase I footprinting. On oocyte chromatin, where the genes are active, the cleavage preferences over the entire gene region showed a periodic pattern of sensitivity and were dramatically different from the patterns obtained with deproteinized DNA or somatic cell chromatin. Further, the normal binding site for TFIIIA over the internal promoter region was preferentially sensitive to cleavage, indicating that TFIIIA was not bound inthe manner predicted by in vitro experiments. In somatic cell chromatin, the oocyte-type 5S genes displayed a cleavage pattern largely similar to deproteinized DNA suggesting the absence of positioned nucleosomes on these inactive genes, although the presence of uncharacterized repressor complexes could not be ruled out. These data are discussed in terms of potential forms of the chromatin structure and alternative mechanisms of oocyte-type gene activation.


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