Supplementary Figure 1 from NUP98 Fusion Oncoproteins Promote Aneuploidy by Attenuating the Mitotic Spindle Checkpoint

Salsi, Valentina; Ferrari, Silvia; Gorello, Paolo; Fantini, Sebastian; Chiavolelli, Francesca; Mecucci, Cristina; Zappavigna, Vincenzo

doi:10.1158/0008-5472.22402251.v1

00085472can130912-sup-fig_1.pdf (2.24 MB)

Supplementary Figure 1 from NUP98 Fusion Oncoproteins Promote Aneuploidy by Attenuating the Mitotic Spindle Checkpoint

journal contribution

posted on 2023-03-30, 22:32 authored by Valentina Salsi, Silvia Ferrari, Paolo Gorello, Sebastian Fantini, Francesca Chiavolelli, Cristina Mecucci, Vincenzo Zappavigna

PDF - 2289K, Supplemental Figure 1. Expression of NUP98 and of NUP98 fusion oncoproteins in HEK293 cells. A, Schematic representation of the structures of human NUP98 (AAC50366) and of its chimeric oncoproteins. The GLFG repeats and the Rae1-interacting GLEBS domain are shown as black and dark grey boxes, respectively. The N-terminal portion of NUP98 is similar in the studied chimeras, spanning aa 1 to aa 469 in NUP98-HOXD13, and aa 1 to aa 514 in NLOC and NHHEX fusions. The HOXD13 (aa 252 to 335, AAC51635), LOC348801 isoform1 (aa 1 to 178, AAI26363), and HHEX (aa 140 to 270, NP_002720) C-terminal moieties are shown as coloured boxes. The homeodomain regions (HD) of HOXD13 and HHEX are indicated as a grey box within the coloured one. Also shown is the structure of NUP98-HOXD13IQN, which carries mutations of three amino acids within the homeodomain (white), impairing DNA binding. B, Western blot analysis showing the expression levels of the indicated proteins transiently expressed in HEK293 cells (T0), and after Nocodazole treatment at the indicated time points. Immunoblottings were performed with anti-NUP98 and anti-betaactin antibodies as a loading control.

History

ARTICLE ABSTRACT

NUP98 is a recurrent fusion partner in chromosome translocations that cause acute myelogenous leukemia. NUP98, a nucleoporin, and its interaction partner Rae1, have been implicated in the control of chromosome segregation, but their mechanistic contributions to tumorigenesis have been unclear. Here, we show that expression of NUP98 fusion oncoproteins causes mitotic spindle defects and chromosome missegregation, correlating with the capability of NUP98 fusions to cause premature securin degradation and slippage from an unsatisfied spindle assembly checkpoint (SAC). NUP98 fusions, unlike wild-type NUP98, were found to physically interact with the anaphase promoting complex/cyclosome (APC/C)Cdc20 and to displace the BubR1 SAC component, suggesting a possible mechanistic basis for their interference with SAC function. In addition, NUP98 oncoproteins displayed a prolonged half-life in cells. We found that NUP98 stability is controlled by a PEST sequence, absent in NUP98 oncoproteins, whose deletion reproduced the aberrant SAC-interfering activity of NUP98 oncoproteins. Together, our findings suggest that NUP98 oncoproteins predispose myeloid cells to oncogenic transformation or malignant progression by promoting whole chromosome instability. Cancer Res; 74(4); 1079–90. ©2013 AACR.