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Nucleic Acids Research, 1990, Vol. 18, No. 16 4825-4831
© 1990

MOLECULAR BIOLOGY

Structural features of the priming signal recognized by primase: mutational analysis of the phage G4 origin of complementary DNA strand synthesis

Hiroshi Hiasa1,2, Hiroshi Sakai1, Tohru Komano1,* and G. Nigel Godson2

1Laboratory of Biochemistry, Department of Agricultural Chemistry, Kyoto University Kyoto 606, Japan 2Department of Biochemistry, New York University Medical Center 550 First Avenue, New York, NY 10016, USA

*To whom correspondence should be addressed

Received April 19, 1990. Revised July 19, 1990. Accepted July 19, 1990.

45 mutations (insertion, deletion and base substitution) of the G4 Goric were tested for their functional activity in M13 and R199 in vivo. The critical mutants were also assayed for their ability to synthesize pRNA in vitro using SSB and primase. The results demonstrate that the secondary structure and spacing of stem-loops I and III are essential for Goric activity and that the 5'-CTG-3' sequence flanking stem-loop I is essential for initiation of pRNA synthesis.


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