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Nucleic Acids Research, 1989, Vol. 17, No. 20 8171-8184
© 1989

MOLECULAR BIOLOGY

Nuclear factors specifically bind to upstream sequences of a Xenopus laevis ribosomal protein gene promoter

Francesca Carnevali*, Claudia La Porta, Vincenza Ilardi and Elena Beccari

Centro di Studio per gli Acidi Nucleici, Department of Genetics and Molecular Biology, University of Rome La Sapienza, P. le A.Moro 5, 00185 Rome, Italy

*To whom correspondence should be addressed

Received June 23, 1989. Revised September 15, 1989. Accepted September 15, 1989.

The upstream region of the Xenopus laevis L14 ribosomal protein gene was deleted starting from the 5' extremity in order to define the promoter length necessary to express a linked reporter CAT gene. The functional analysis indicated that a sequence located between –63 and –49 from the capsite is important for an efficient promoter activity. Band shift and ExoIII protection assays evidenced the binding to this region of a factor, called XrpFI, present in the crude nuclear extract from X. laevis oocytes. Methylation interference analysis localized the contacts in the G residues belonging to a short box, 5' CTTCC 3', positioned between –53 and –49 from the capsite. An additional factor, XrpFII, makes contacts with the sequence 5'GCCTGTTCGCC 3' located between –27 and –17 from the capsite. The deletion mutant still containing this sequence is poorly transcribed, but resumes activity when a short fragment containing the binding site for factor XrpFI is cloned in an upstream position.


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