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. 1998 Jan;18(1):420–432. doi: 10.1128/mcb.18.1.420

FIG. 3.

FIG. 3

Effects of deletions within the 81-amino-acid domain on activity of TFIIIA. (A) Amino acid sequences of zinc finger 8, the 81-amino-acid domain, and zinc finger 9 of yeast TFIIIA, as shown in Fig. 1B. Rectangles and numbers above the amino acid sequence represent regions deleted from the 81-amino-acid domain. The deletions in TFIIIA(Δ284-364), which lacks the entire 81-amino-acid domain, and in TFIIIA(Δ282-353) are not represented. (B) Abilities of versions of TFIIIA with deletions within the 81-amino acid-domain to bind the 5S RNA gene and to recruit TFIIIC to the TFIIIA-DNA complex, as outlined in the legend for Fig. 1C. Lanes 1 to 4 were as described for Fig. 1C. Lanes 5 to 20, in vitro-synthesized versions of TFIIIA lacking residues 284 to 364 (lanes 5 and 6), 282 to 287 (lanes 7 and 8), 282 to 315 (lanes 9 and 10), 282 to 353 (lanes 11 and 12), 316 to 334 (lanes 13 and 14), 328 to 341 (lanes 15 and 16), 342 to 351 (lanes 17 and 18), or 352 to 359 (lanes 19 and 20). (C) Abilities of versions of TFIIIA with deletions in the 81-amino-acid domain to support in vitro transcription of the 5S RNA gene. For details, see the legend to Fig. 1D. (D) Abilities of versions of TFIIIA with deletions in the 81-amino-acid domain to support cell viability. A plasmid shuffle system was used to test the abilities of mutant versions of TFIIIA to support cell viability as described in the legend to Fig. 2D. Growth of cells containing pG3-derived plasmids on medium containing 5-FOA is shown. vector, pG3 not expressing TFIIIA; WT, pG3 expressing wild-type TFIIIA; Δn-n, pG3 expressing a version of TFIIIA that lacks amino acids n to n. (E) Assessment by Western blot analysis of in vivo expression of TFIIIA containing deletions in the 81-amino-acid domain. The analysis was carried out as described in the legend to Fig. 2E. Protein in lanes 1 to 9 was extracted from YRW1 cells containing pG3-derived plasmids expressing no TFIIIA, wild-type TFIIIA, TFIIIA(Δ284-364), TFIIIA(Δ282-315), TFIIIA(Δ282-353), TFIIIA(Δ316-334), TFIIIA(Δ328-341), TFIIIA(Δ342-351), and TFIIIA(Δ352-359), respectively. The asterisk on the left indicates a cross-reacting molecule not related to TFIIIA. Only the portion of the blot that contains TFIIIA is shown.