A conserved CENP-E region mediates BubR1-independent recruitment to the outer corona at mitotic onset.

The outer corona plays an essential role at the onset of mitosis by expanding to maximize microtubule attachment to kinetochores.1,2 The low-density structure of the corona forms through the expansion of unattached kinetochores. It comprises the RZZ complex, the dynein adaptor Spindly, the plus-end directed microtubule motor centromere protein E (CENP-E), and the Mad1/Mad2 spindle-assembly checkpoint proteins.3,4,5,6,7,8,9,10 CENP-E specifically associates with unattached kinetochores to facilitate chromosome congression,11,12,13,14,15,16 interacting with BubR1 at the kinetochore through its C-terminal region (2091-2358).17,18,19,20,21 We recently showed that CENP-E recruitment to BubR1 at the kinetochores is both rapid and essential for correct chromosome alignment. However, CENP-E is also recruited to the outer corona by a second, slower pathway that is currently undefined.19 Here, we show that BubR1-independent localization of CENP-E is mediated by a conserved loop that is essential for outer-corona targeting. We provide a structural model of the entire CENP-E kinetochore-targeting domain combining X-ray crystallography and Alphafold2. We reveal that maximal recruitment of CENP-E to unattached kinetochores critically depends on BubR1 and the outer corona, including dynein. Ectopic expression of the CENP-E C-terminal domain recruits the RZZ complex, Mad1, and Spindly, and prevents kinetochore biorientation in cells. We propose that BubR1-recruited CENP-E, in addition to its essential role in chromosome alignment to the metaphase plate, contributes to the recruitment of outer corona proteins through interactions with the CENP-E corona-targeting domain to facilitate the rapid capture of microtubules for efficient chromosome alignment and mitotic progression.

Circumcaval ureter associated with an intrahepatic portosystemic shunt in a dog.

CASE DESCRIPTION: A 4-month-old Bernese Mountain Dog was examined because of shifting hind limb lameness and lethargy of 2 weeks' duration. CLINICAL FINDINGS: The lameness was attributed to hypertrophic osteodystrophy. Portosystemic shunting was suspected on the basis of low serum albumin concentration and high serum bile acids concentration, and an intrahepatic shunt was identified ultrasono-graphically. Celiotomy was performed, and the shunt was partially closed with a cellophane band. During follow-up ultrasonography 7 months later, dilation of the left renal pelvis and proximal portion of the left ureter was identified. During exploratory celiotomy, the left ureter was found to pass dorsal to the caudal vena cava, and circumcaval ureter was diagnosed. TREATMENT AND OUTCOME: The ureter was transected, repositioned ventral to the vena cava, and anastomosed. Follow-up ultrasonographic examinations revealed gradual resolution of the hydronephrosis and hydroureter. CLINICAL RELEVANCE: Findings suggest that circumcaval ureter should be considered in the differential diagnosis for hydronephrosis and hydroureter in dogs. Partial obstruction of the middle segment of the ureter on ultrasonograms or contrast radiographs should increase the index of suspicion for this condition.