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. 2013 Nov 6;288(51):36676–36690. doi: 10.1074/jbc.M113.508267

FIGURE 5.

FIGURE 5.

Internal NN cluster mediates premature termination by responding to Nrd1(CIDRtt103) at the CRH1 gene. A, an internal coding region of CRH1 containing the NN cluster was replaced with a same length of DNA fragment from EGFP. B, CRH1 transcripts derived from either wild-type CRH1 or the CRH1::EGFP gene were analyzed by Northern blotting upon depletion of Rrp41. The relative portion of premature-terminated short transcripts (marked by green arrowhead) was quantitated as described in the legend in Fig. 4C. Shorter transcripts were significantly diminished in Nrd1(CIDRtt103) Δrtt103 cells when the NN cluster was removed. C, schematic diagram of CRH1 with internal NN clusters. Positions of ChIP PCR primers are shown above the diagram. D, RNApII occupancy was significantly decreased near the internal NN cluster (position 4) in Nrd1(CIDRtt103) Δrtt103 cells when compared with WT. RNApII occupancy signals at each position from WT were set to 100 and those from Nrd1(CIDRtt103) Δrtt103 cells were calculated in relative amounts after normalization to a non-transcribed telomeric region. Error bars represent S.E. for three independent experiments.