Supplementary Information for Telomeric RNA-DNA hybrids affect telomere length dynamics and senescence Bettina Balk 1,2 , André Maicher 1,2 , Martina Dees 1 , Julia Klermund 1 , Sarah Luke- Glaser 1 , Katharina Bender 1 and Brian Luke 1,3 1 Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ- ZMBH Alliance, Heidelberg, Germany 2 These authors contributed equally List of Supplementary Information Supplementary Figures (5) Supplementary Tables (3) Nature Structural and Molecular Biology: doi:10.1038/nsmb.2662
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Supplementary Information for - Nature Research · 2013-10-04 · calculated (est2 n = 48, PD 30; est2 hpr1 n = 39; PD 26). a b Relative cell density est2 rad52 est2 rad52 hpr1 Average
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Supplementary Information
for
Telomeric RNA-DNA hybrids affect telomere length dynamics and senescence
Bettina Balk1,2, André Maicher1,2, Martina Dees1, Julia Klermund1, Sarah Luke-
Glaser1, Katharina Bender1 and Brian Luke1,3
1Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ-
ZMBH Alliance, Heidelberg, Germany
2These authors contributed equally
List of Supplementary Information
Supplementary Figures (5)
Supplementary Tables (3)
Nature Structural and Molecular Biology: doi:10.1038/nsmb.2662
Supplementary Figure 1. Telomeric RNA-DNA hybrids are regulated by
RNase H enzymes
(a) RNA-DNA hybrids exist at telomeres and accumulate in RNase H mutants. The
identical ChIP as described in Fig. 1a, however quantified showing % input on the Y-
axis. Values are represented as % input of telomeric DNA recovered and are the
means of seven (+ab) and five (-ab) biological replicates, error bars depict ± s.e.m.
P values were derived from two-tailed Student’s t-tests (NS = not significant). (b) The
RNA-DNA hybrid signal is RNase H sensitive. Following overnight
immunoprecipitation with the S9.6 antibodies, one wild type sample was treated with
recombinant RNase H before the washing steps (see online methods). (c) TERRA
levels are not increased in rnh1 rnh201 mutants. qRT-PCR was performed for the
6Y’, 1L and 15L telomeres using the indicated mutants (at approx. PD 15 following
tetrad dissection). sir2 cells served as positive control where TERRA is upregulated.
Average values were derived from 3 biological replicates where error bars depict ±
s.e.m.
ba
c
WT
rnh1 rnh201
0
0.2
0.4
0.6
0.8
1.0
ab
RNase H
6Y´15L rDNA
+ +
+
1L
0
0.05
0.10
0.60
0.80
1.00
P = 0.46
ab + + +
6Y´15L rDNA
+
1L
NS
P = 0.61
P = 0.001
***
P = 0.03
*P = 0.003
**
NS
P = 0.09
**P =0.005
NS
P = 0.02
*
% I
np
ut
% I
np
ut
Fo
ld e
nrich
me
nt
co
mp
are
d t
o W
T
0
100
200
300
1
2
3
4
5
1L 15L
WT
rnh1 rnh201est2est2 rnh1 rnh201sir2
– – – –
–
– –
+ +
+
–
– –
+ +
+
–
– –
+ +
+
–
– –
Nature Structural and Molecular Biology: doi:10.1038/nsmb.2662