Centromeric H2B monoubiquitination promotes noncoding transcription and chromatin integrity

Functional centromeres are essential for proper cell division. Centromeres are established largely by epigenetic processes resulting in incorporation of the histone H3 variant CENP-A. Here, we demonstrate the direct involvement of H2B monoubiquitination, mediated by RNF20 in humans or Brl1 in Schizosaccharomyces pombe, in centromeric chromatin maintenance. Monoubiquinated H2B (H2Bub1) is needed for this maintenance, promoting noncoding transcription, centromere integrity and accurate chromosomal segregation. A transient pulse of centromeric H2Bub1 leads to RNA polymerase II–mediated transcription of the centromere’s central domain, coupled to decreased H3 stability. H2Bub1-deficient cells have centromere cores that, despite their intact centromeric heterochromatin barriers, exhibit characteristics of heterochromatin, such as silencing histone modifications, reduced nucleosome turnover and reduced levels of transcription. In the H2Bub1-deficient cells, centromere functionality is hampered, thus resulting in unequal chromosome segregation. Therefore, centromeric H2Bub1 is essential for maintaining active centromeric chromatin. ChIP: In total 17 samples; 15 ChIP DNA files (7 different conditions) 2 Input files. Cnp1_WT_input.bar (antibody against Cnp1 in strains Hu303, Hu29 (WT) (Cnp1_Hu303.CEL, AS_10.CEL) vs input (JW_1.CEL, JW_2_2.CEL)); Cnp1_Hu2640_WT.bar (antibody against Cnp1 in strain Hu2640 (htb1K119R) (Cnp1_Hu2640_A.CEL, Cnp1_Hu2640_B.CEL) vs strains Hu303, Hu29 (WT) (Cnp1_Hu303.CEL, AS_10.CEL)); Cnp1_Hu2640_input.bar (antibody against Cnp1 in strain Hu2640 (htb1K119R) (Cnp1_Hu2640_A.CEL, Cnp1_Hu2640_B.CEL) vs input (JW_1.CEL, JW_2_2.CEL)); H2Bub1_Hu303_input.bar (antibody against H2Bub1 in strain Hu303 (WT) (H2Bub1_Hu303_A.CEL, H2Bub1_Hu303_B.CEL) vs input (JW_1.CEL, JW_2_2.CEL)); H3_WT_input.bar (antibody against H3 in strains Hu303, Hu29 (WT) (H3_Hu303.CEL, As_12.CEL) vs input (JW_1.CEL, JW_2_2.CEL); H3_Hu2640_WT.bar (antibody against H3 in strain Hu2640 (htb1K119R) (H3_Hu2640_A.CEL, H3_Hu2640_B.CEL, H3_Hu2640_C.CEL) vs strains Hu303, Hu29 (WT) (H3_Hu303.CEL, AS_12.CEL)); H3_Hu2640_input.bar (antibody against H3 in strain Hu2640 (htb1K119R) (H3_Hu2640_A.CEL, H3_Hu2640_B.CEL, H3_Hu2640_C.CEL) vs input (JW_1.CEL, JW_2_2.CEL)); H3K9me2_Hu303_input.bar (antibody against H3K9me2 in strains Hu303, Hu29 (WT) (H3K9me2_Hu303.CEL, AS_6.CEL) vs input (JW_1.CEL, JW_2_2.CEL)); H3k9me2_Hu2640_Hu303.bar (antibody against H3K9me2 in strain Hu2640 (htb1K119R) (H3K9me2_Hu2640_A.CEL, H3K9me2_Hu2640_B.CEL) vs strain Hu303, Hu29 (WT) (H3K9me2_Hu303.CEL, AS_6.CEL)); H3K9me2_Hu2640_input.bar (antibody against H3K9me2 in strain Hu2640 (htb1K119R) (H3K9me2_Hu2640_A.CEL, H3K9me2_Hu2640_B.CEL) vs input (JW_1.CEL, JW_2_2.CEL)). RNA: 4 RNA samples: 2 replicates of WT RNA (RNA_Hu303_A.CEL, RNA_Hu303_B.CEL) and 2 replicates of htb1-K119R RNA (RNA_Hu2640_A.CEL, RNA_Hu2640_B.CEL).