Extended Data Fig. 9: Transcription at pericentromere borders influences cohesin position.
a, Genes at pericentromere borders are moderately transcribed on average. Shown is the relative RNA density for convergent border gene pairs compared with all genes under conditions of no tension or tension (n = 1). RNA-seq analysis of wild-type cells arrested in metaphase in the presence or absence of tension. Dashed lines indicate means; dotted lines mark 95% confidence intervals. b, Box plot showing transcription levels of genes at pericentromere borders based on RNA polymerase II (Rpo21) cross-linking and analysis of cDNA (CRAC) from ref. 29. Rpo21 CRAC sense read counts of genes at borders were normalized to the protein-coding-gene average, and genes at pericentromere borders were grouped by their relative orientation to centromeres. Data points correspond to means of three biological repeats. Centre lines, medians; box limits, second and third quartiles; whiskers, first and fourth quartiles (non-normal distribution, Shapiro–Wilk; *P < 0.05, two-sided Mann–Whitney test). c, Box plot showing relative transcription levels of genes transcribed towards and away from centromeres, at pericentromere borders and at non-border convergent genes inside pericentromeres, as in b. d, e, Insertion of a URA3 cassette between a convergent gene pair shifts the localization of cohesin in the direction of transcription. URA3 was integrated in either orientation between the convergent gene pairs at the left pericentromere border on chromosome IV, and cohesin (Scc1) ChIP–qPCR (n = 3; bars show means ± s.e.m.) using primers at the indicated positions (d) and ChIP-seq (n = 1) (e) were performed.
