NPL4 function is required for
OLE1 transcription. Northern analysis was performed on
total RNA isolated from ole1Δ (lane 1), wild-type (WT;
lanes 2–4), npl4-1 (lanes 5–7), npl4-2
(lanes 8–10), cdc48-3 (lanes 11–13),
ufd1-1 (lanes 14-16), and rpn4Δ cells
(lanes 17–19) with OLE1- and
ACT1-specific DNA probes. Cells were grown in YEPD
media at 25°C with or without supplemented UFAs. For wild-type cells,
the temperature-sensitive-strains npl4-1,
npl4-2, and cdc48-3, and
non–temperature sensitive strains ufd1-1 and
rpn4Δ, samples were also collected after shifts to
37°C for 60 min. A normalized OLE1/ACT1
ratio was calculated (see MATERIALS AND METHODS) for each sample to
allow for quantitation of the relative changes in OLE1
expression among samples.