TY - JOUR
T1 - Identification and sequence of the initiation slte for rat 45S ribosomal RNA synthesis
AU - Harrington, Christina A.
AU - Chikaraishi, Dona M.
N1 - Funding Information:
ACKNOWLEDGEMENTS We thank Barbara Masters and James Beeson for Identifying and Isolating lambda clone 1A. We are Indebted to Lisa Buchanan for technical assistance and to Cathy Inouye and Lynn Van Meter for typing of the manuscript and Figure 4. We thank Drs. Elaine Lewis and Thomas Walker for helpful discussion and critical reading of the manuscript. This work was supported by Grant GM 29302 from the National Institute of Health. C.A.H. was supported by NIH grant 5 T32 GM 07385-04 from 1/81 to 7/81.
PY - 1983/5/25
Y1 - 1983/5/25
N2 - 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.
AB - 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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U2 - 10.1093/nar/11.10.3317
DO - 10.1093/nar/11.10.3317
M3 - Article
C2 - 6304628
AN - SCOPUS:0021111958
SN - 0305-1048
VL - 11
SP - 3317
EP - 3332
JO - Nucleic acids research
JF - Nucleic acids research
IS - 10
ER -