Extended Data Fig. 7: The multi-phase separation of nucleoids is remodeled by TFB2M in promoter melting.
From: Phase separation drives the self-assembly of mitochondrial nucleoids for transcriptional modulation
a, The multi-phase separation of transcription initiation complex after promoter melting. The paired two strand 52 bp LSP promoter DNA (LSP-WT) or a melted LSP DNA (LSP-melt) containing a 7 bp unpaired sequence was used in droplet formation with transcription initiation complex. All the sample contains 200 nM DNA, 20 µM TFAM and 100 nM POLRMT. Mock, without TFB2M and ATP; + TFB2M, add TFB2M; + TFB2M + ATP, add TFB2M and 200 µM ATP in phase separation. Scale bar, 5 μm. b, The percentage of layered structure of POLRMT of all droplets was counted in left images (a). c, The multi-phase separation of transcription initiation complex after promoter melting. The paired two strand 50 bp HSP promoter DNA (HSP-WT) or a 50 bp melted HSP DNA (HSP-melt) containing a 7 bp unpaired sequence was used in transcription initiation complex droplet formation. All the sample contains DNA, TFAM and POLRMT. Mock, without TFB2M and ATP; + TFB2M, add TFB2M; + TFB2M + ATP, add TFB2M and 200 µM ATP in phase separation. Scale bar, 5 μm d, The percentage of layered structure of POLRMT of all droplets was counted in left images (c). Data in (b), (d) are present with mean ± s.d. e,f, The mitochondrial transcription level after the different dose overexpression of TFB1M (e) or TFB2M (f). Data are present with mean ± s.d., and n = 3 independent experiments. Two-tailed unpaired Student’s t-test was used. The p values were noted on the panel. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.
