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Nucleic Acids Research, 1989, Vol. 17, No. 4 1605-1618
© 1989

ENZYMOLOGY

Abortive initiation by bacteriophage T3 and T7 RNA polymerases under conditions of limiting substrate

Mei-Ling Ling1, Steven S. Risman, John F. Klement1, Nancy McGraw1 and William T. McAllister

Department of Microbiology and Immunology, Morse Institute for Molecular Genetics, New York Univesity, Brooklyn Health Science Center 450 Clarkson Avenue, Box 44, Brooklyn, NY 11203-2098 1Department of Microbiology, Rutgers Medical School Piscataway, NJ 08854, USA

Received October 31, 1988. Accepted January 6, 1989.

Initiation of RNA synthesis by the phage polymerases is atortive if the concentration of pyrimidine triphosphates is limiting. Under abortive initiation conditions the polymerases repeatedly initiate transcription but produce ribooligonucleotides that terminate just prior to the first occurence of the limiting substrate. Abortive initiation is most severe if the limiting substrate occurs within the first 8–12 nucleotides of the nascent RNA chain and is particularly evident when UMP is limiting. The formation of stable elongation complexes (as determined by gel retardation experiments) occurs after the synthesis of an RNA product 8–12 nucleotides in length.


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