Impaired CENP-E Function Renders Large Chromosomes More Vulnerable to Congression Failure

Abstract

It has recently emerged that human chromosomes vary between one another in terms of features that impact their behaviour during impaired chromosome segregation, leading to non-random aneuploidy in the daughter cell population. During the process of chromosome congression to the metaphase plate, chromosome movement is guided by kinesin-like proteins, among which centromere-associated protein E (CENP-E) is important to transport chromosomes along the microtubules of the mitotic spindle. It is known that the inhibition of CENP-E notably impairs alignment for a subset of chromosomes, particularly those positioned close to the centrosome at nuclear envelope breakdown (‘polar chromosomes’); it is, however, not clear whether chromosome identity could influence this process. Since a popular strategy to model aneuploidy is to induce congression defects (for example combining CENP-E inhibitors with mitotic checkpoint abrogation), variance in congression efficiency between chromosomes might influence the landscape of aneuploidy and subsequent cell fates. By combining immunofluorescence, live cell imaging and fluorescence in situ hybridisation, we investigated the behaviour of polar chromosomes and their dependency upon CENP-E-mediated congression in human cells. We observed a bias in congression efficiency related to chromosome size, with larger chromosomes more sensitive to CENP-E inhibition. This bias is likely due to two contributing factors; an initial propensity of larger chromosomes to be peripheral and thus rely more upon CENP-E function to migrate to the metaphase plate, and additionally a bias between specific chromosomes’ ability to congress from a polar state. These findings may help to explain the persistence of a subset of chromosomes at the centrosome following CENP-E disruption, and also have implications for the spectrum of aneuploidy generated following treatments to manipulate CENP-E function.

Human chromosome‐specific aneuploidy is influenced by DNA ‐dependent centromeric features

“…Surprisingly, despite these features being mainly dependent on centromere length, the size of HOR arrays per se does not seem to directly influence segregation fidelity. It has been previously observed that larger chromosomes tend to mis‐segregate more frequently in cancer cells (Bochtler et al , ) or under conditions that compromise the function of CENP‐E (Tovini & McClelland, ), a key component of chromosome congression (Kapoor et al , ). Here, we also found that larger chromosomes have a slightly higher tendency to mis‐segregate, but this correlation is strongly enhanced by the diversity in the centromere features (centromere strength) over the whole chromosome length (Fig A and B).…”

The protective effect and mechanism of catalpol on high glucose-induced podocyte injury

“…Bax, caspase-3, Bcl-2 and NF-κB levels were determined by immunofluorescence staining, which was performed according to methods described previously [19]. Briefly, after blocking with 5% BSA, podocytes were incubated with primary antibodies against Bax, caspase-3, Bcl-2 and NF-κB diluted 1:100.…”

Polar Chromosomes—Challenges of a Risky Path

“…These findings imply that only a low number of unattached polar chromosomes could induce moderate mitotic delays, which would lead to mis-segregation of unaligned chromosomes. Accordingly, studies that used CENP-E perturbations reported mis-segregation of one or a few unaligned polar chromosomes after moderate mitotic delays in HeLa [103], RPE1 [188,189], and mouse embryonic cells [190]. A similar phenomenon was observed after treatment of cells with nanomolar doses of nocodazole [186].…”