Centromeric Transcription Regulates Aurora-B Localization and Activation

Abstract

Summary Centromeric transcription is widely conserved, however it is not clear what role centromere transcription plays during mitosis. Here I find that centromeres are transcribed in Xenopus egg extracts into a long noncoding RNA (lncRNA; cen-RNA) that localizes to mitotic centromeres, chromatin, and spindles. Cen-RNAs bind to the Chromosomal Passenger Complex (CPC) in vitro and in vivo. Blocking transcription or antisense inhibition of cen-RNA leads to a reduction of CPC localization to the inner centromere and misregulation of CPC component Aurora-B activation, independently of known centromere recruitment pathways. Additionally, transcription is required for normal bipolar attachment of kinetochores to the mitotic spindle, consistent with a role for cen-RNA in CPC regulation. This work demonstrates that cen-RNAs promote normal kinetochore function through regulation of the localization and activation of the CPC and confirm that lncRNAs are components of the centromere.

Human Centromeres Produce Chromosome-Specific and Array-Specific Alpha Satellite Transcripts that Are Complexed with CENP-A and CENP-C

“…Our IF-RNA FISH results suggest that only a fraction of alpha satellite transcripts co-localizes with centromeric chromatin. These findings agree with previous studies reporting that non-coding centromeric RNAs in various organisms are present at the inner centromere (Blower, 2016; Ideue et al, 2014; Liu et al, 2015; Wong et al, 2007). …”

“…The identity of the RNA polymerase responsible for centromeric transcription has been an open area of debate, because evidence exists for the involvement of RNA polymerases (RNAP) I, II, and III (Blower, 2016; Chan et al, 2012; Grenfell et al, 2016; Liu et al, 2015; Longo et al, 2015; Quenet and Dalal, 2014; Wong et al, 2007). To identify the polymerase that transcribes the chromosome- and array-specific alpha satellite transcripts, we pharmacologically inhibited RNAPs.…”

“…Transcription is an important requirement of centromere assembly in various eukaryotes (Blower, 2016; Carone et al, 2009; Chan et al, 2012; Chen et al, 2015; Grenfell et al, 2016; Lu and Gilbert, 2007; Molina et al, 2016; Rosic et al, 2014; Wong et al, 2007). Our study also emphasizes that alpha satellite transcription is a key feature of eukaryotic centromeres.…”

“…Studies in various organisms have demonstrated that transcription occurs within centromeres and is involved in CENP-A loading and kinetochore assembly (Blower, 2016; Catania et al, 2015; Grenfell et al, 2016; Liu et al, 2015; Quenet and Dalal, 2014; Rosic et al, 2014; Topp et al, 2004; Wong et al, 2007). In humans, RNA polymerase II (RNAP II) has been observed at centromeres in G1 and at mitosis (Quenet and Dalal, 2014; Wong et al, 2007), implicating transcription in centromere function.…”

The chromosomal basis of meiotic acentrosomal spindle assembly and function in oocytes

“…There could be epigenetic factors such as histone modifications (Kelly, et al 2010; Wang, et al 2010; Yamagishi, et al 2010) or proteins like HP1 (Abe, et al 2016; Ainsztein, et al 1998), which are known to recruit the CPC. An interesting recent development is that noncoding RNAs may have a role in chromatin-mediated spindle assembly (Blower 2016; Chen, et al 2015; Grenfell, et al 2016; Jambhekar, et al 2014). …”