2 FORMS OF REPLICATION INITIATOR PROTEIN - POSITIVE AND NEGATIVE CONTROLS

The pir gene of plasmid R6K encodes the protein, pi, a replication and transcription factor. Two translational options for the pir gene give rise to two forms of pi protein: a 35.0-kDa form (pi(35.0)) and a sho rtened 30.5-kDa form (pi(30.5)). Although both proteins bind to a seri es of 22-bp direct repeats essential for plasmid R6K replication, only pi(35.0) can bind to a site in the (A.T)-rich segment of its y ori an d activate the gamma ori in vivo and in vitro. However, unlike pi(35.0 ), pi(30.5) can inhibit in vivo and in vitro replication (activated by pi(35.0)). We propose that the two forms of pi might have distinct fu nctions in replication. We show that although both forms of pi produce dimers, the nature of these dimers is not identical. The N-terminal 3 7 amino acid residues appear to control the formation of the more stab le pi(35.0) dimers, whereas another, apparently weaker interface holds together dimers of pi(35.0) . We speculate that the leucine zipper-li ke motif, absent in pi(30.5), controls very specific functions of pi p rotein.