SRG1 is a promoter associated RNA whose decay requires Rrp6p.
(A) Northern blot to detect the levels of
poly(A)+ and poly(A)− SRG1 RNA (Upper):
3 μg of total RNA from wild type,
trf4Δ, and rrp6Δ
(lanes 1, 4, and 7), 100 ng of poly(A)+ RNA from wild type,
trf4Δ, and rrp6Δ
(lanes 2, 5, and 8), and 3 μg of poly(A)− RNA from wild
type, trf4Δ, and rrp6Δ
(lanes 3, 6, and 9). The membrane was probed with an SRG1-specific
oligo. (Lower) A duplicate membrane probed with an
oligo complementary to snR128. (B) Northern blot
analysis of SER3 in different yeast mutants. Three
micrograms of total RNA from wild-type (lane 1),
trf4Δ (lane 2), rrp6Δ
(lane 3), heat-shifted spt5-194
(lane 4), heat-shifted spt5-194,
rrp6Δ (lane 5), and
snf2Δ (lane 6) yeast were fractioned on a
1% agarose formaldehyde gel. The membrane was probed with an
SRG1+SER3 oligo complementary to a region contained in both
SRG1 RNA and SER3
mRNA.