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Nucleic Acids Research, 1983, Vol. 11, No. 10 3317-3332
© 1983

MOLECULAR BIOLOGY

Identification and sequence of the initiation slte for rat 45S ribosomal RNA synthesis

Christina A. Harrington* and Dona M. Chikaraishi

Division of Molecular and Cellular Biology, National Jewish Hospital and Research Center National Asthma Center Denver Co, 80206 *Department of Molecular Cellular and Developmental Biology, University of Colorado Boulder, Co 80309, USA

Received October 25, 1983. Revised April 6, 1983. Accepted April 6, 1983.

The transcription Initiation site for rat 45S precursor ribosomal RNA synthesis was determined by nuclease protection mapping with two single-strand endonucleases, SI and mung bean, and one single-strand exonuclease, ExoVII. These experiments were performed with end-labeled ribosomal DNA from double-stranded pBR322 recombinants and from single-stranded M13 recombinants. Results from experiments using both kinds of DNA and all three enzymes showed that the 5' end of 45S RNA mapped to a unique site 125 bases upstream from the H1nd III site 1n the ribosoraal DNA gene. The DNA surrounding this site (designated +1) was sequenced from -281 to +641. The entire sequence of this region shows extensive homology to the comparable region of mouse. This Includes three stretches of T residues 1n the non-coding strand between +300 and +630. Two sets of direct repeats adjacent to these T-r1ch regions are observed. Comparison of the mouse and human ribosomal DNA transcription Initiation sites with the rat sequence reported 1n this paper demonstrates a conserved sequence at +2 to +16, CTGACACGCTGTCCT. This suggests that this region may be Important for the Initiation of transcription on mammalian ribosomal UNAs


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