The tL2 cluster of transcription termination sites between genes bet and ral of coliphage lambda

The major leftward transcription of bacteriophage lambda is controlled by several terminators (t), including tL1, tL2, tL3, and others. The tL2 termination site, which was placed by Salstrom and Szybalski (Virology 88, 252-260, 1978) between lambda genes bet and ral, was found to consist of a cluster of four leftward terminators. As not to change the numbering of other leftward terminators, these were designated as tL2a, tL2b, tL2c, and tL2d. As determined by S1 nuclease mapping of the tL2-terminated in vitro transcripts, the normally pL-initiated major leftward lambda transcription should encounter termination points at 1653 bp (tL2a; between genes Ea10 and ral), 2089 bp (tL2b; between genes cIII and Ea10), 2441-2442 bp, and 2483 bp (tL2c and tL2d; both within gene gam) from the sL startpoint (= +1). All terminators were cloned in a pBR322-derived plasmid between the p'R promoter and the galK gene, and their in vivo termination efficiencies are 69% (tL2a), 53% (tL2b), and 38% (tL2c + tL2d), measured as reduction of galK expression in rho+galK- hosts at 30 degrees. The tL2a and tL2b), terminators depend little on the rho factor, whereas the efficiency of tL2c + tL2d decreases from 38 to only 14% in the rho- host. When shifted to 42 degrees, the termination efficiency of tL2b decreases from 53 to only 36%, while the other tL2 terminators are much less affected by increasing the temperature. The calculated joint efficiency of the entire tL2 cluster is 90%, which is in perfect agreement with the 90% termination efficiency reported by Salstrom and Szybalski (1978) for tL2. However, the natural location of tL2c and tL2d within the actively translated gam gene may interfere with their termination function. Under in vitro conditions, tL2c and tL2d are active only in the presence of rho factor, whereas tL2a and tL2b do not require rho. The structure of the tL2 terminators resembles that of some other known termination sites: a perfect (8 bp for tL2a and tL2b) or imperfect (8-9 bp) dyad symmetry and a T6 (tL2a), T5 (tL2b), T4 (tL2c) or TTATT sequence (tL2d) toward the 3' end of the mRNA-like DNA strand.