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RTP–hSPT5 interactions can reverse guanylylation stimulation. (A) Purified RGT fragment (3.5 pmoles) was incubated with [α-32P]GTP in the presence of 0, 7, 14, and 28 pmoles of hSPT5 and assayed as in Fig. 2B. (B) HCE (3.5 pmoles, lanes 1–5) was assayed for guanylyltransferase activity in the presence of hSPT5 (7 pmoles, lanes 2–5) and 7, 14, or 28 pmoles of MCE–RTP fragment (lanes 3–5). Guanylylation of 32P-labeled HCE was quantitated by PhosphorImager after SDS-PAGE.
