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Nucleic Acids Research, 1989, Vol. 17, No. 9 3459-3468
© 1989

MOLECULAR BIOLOGY

Prohead RNA of bacteriophage {pi}29: size, stoichiometry and biological activity

Jesdawan Wichitwechkarn, Suzanne Bailey1, James W. Bodley1 and Dwight Anderson

Departments of Microbiology/Dentistry Minneapolis, MN 55455, USA 1Departments of Biochemistry, University of Minnesota Minneapolis, MN 55455, USA

Received January 23, 1989. Revised March 27, 1989. Accepted March 27, 1989.

We previously demonstrated (Guo et al., 1987. Nucl. Acids Res. 15, 7081–7090) that purified proheads of bacteriophage {pi}29 contain an RNA of 120 bases which is essential for DNA packaging. Here we report that this RNA exists primarily as a polymer of ca. 174 residues in phage-infected cells and that ca. 54 bases are cleaved from its 3'-terminus by adventitious nucleases during the purification of proheads. The long and short forms of the RNA had similar activity in in vitro DNA packaging and phage assembly. We report the sequence of the long form of the RNA and show that similar long and short forms can be isolated from the proheads of the {pi}29 relatives {varphi}21, {varphi}15 and SF5. The concentration dependence in the reconstitution of RNA-free proheads suggests that one copy of the RNA is sufficient to restore DNA packaging activity to RNA-free proheads. However, quantitative measurements indicate that 5 to 6 copies of the RNA are present on proheads isolated from phage-infected cells.


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