ASH1 mRNA and IST2 mRNA are delocalized in cells expressing She2p mutants N36S and R63K. (A) Strain YLM851 was transformed with plasmids YCplac111 (vector), pRL199 (She2p-wt), pRL322 (She2p-R63K), or pRL324 (She2p-N36S). Transformants were patched on low adenine media devoid of leucine or streaked for single colonies on media devoid of leucine and arginine in the (+) presence and (−) absence of canavanine. (B) Strain YLM777 was transformed with plasmid C3431 (YEplac195/ASH1) and pRL199, pRL322, or pRL324. Transformants were grown in media devoid of leucine and uracil. Cells were processed for in situ hybridization using Cy3-conjugated complementary DNA probes for ASH1 mRNA. Representative images for in situ hybridization, DAPI staining, and (Nom.) Nomarski optics are shown. (C) Strain YLM777 was transformed with plasmid pRL277 [pHZ18(A)-IST2] and pRL199, pRL322, or pRL324. Cells were processed for in situ hybridization using Cy3-conjugated complementary DNA probes for lacZ-IST2 mRNA.
