Rabbit Anti-CDC40 monoclonal antibody, clone TE196-0

Previous work has identified a group of genes whose products play important roles in two seemingly unrelated processes: cell cycle progression and splicing. The products of these genes show a network of physical and genetic interactions suggestive of the existence of a protein complex, the cell cycle and splicing complex (CSC). Here we analyze the genetic interactions between ISY1, SYF2 and NTC20, three non-essential components of the CSC. We show that mutations in ISY1 cause lethality in the absence of Ntc20p, and that the double mutant isy1Delta syf2Delta shows a temperature-dependent cell cycle arrest. This arrest is due to lower levels of alpha-tubulin, a protein encoded by TUB1 and TUB3, two intron-containing genes. We show that the low levels of alpha-tubulin in isy1Delta syf2Delta trigger activation of the spindle checkpoint, causing cell cycle arrest. Thus, our results have uncovered an unexpected role for pre-mRNA splicing in the maintenance of the fidelity of chromosome transmission during cell division.

Simple Summary Paratuberculosis or Johne's disease (JD) in cattle is a chronic granulomatous gastroenteritis caused by infection with Mycobacterium avium subspecies paratuberculosis (MAP). JD is not treatable; therefore, the early identification of infected animals is a key point in reducing its incidence worldwide. In this paper, combinatorial discriminant analysis was applied to transcriptomic data to find a small set of differentially expressed genes able to discriminate between exposed cattle from non-exposed animals. Results of the discriminant analysis identified 10 transcripts that differentiate between ELISA-negative animals belonging to paratuberculosis-positive herds that were potentially exposed to MAP and negative-unexposed animals belonging to paratuberculosis-negative herds. The 10 transcript signature was also used for the classification of 5 cattle positive-for-MAP infection samples, with the underlying hypothesis that their transcriptional profile should be closer to potentially exposed animals than to controls. The same set of 10 transcripts was able to differentiate ELISA-negative unexposed animals from positive animals based on the results of the ELISA test for bovine paratuberculosis and faecal culture. In conclusion, these findings suggest the possible use of the identified RNA expression signature as a new diagnostic test for paratuberculosis. Paratuberculosis or Johne's disease in cattle is a chronic granulomatous gastroenteritis caused by infection with Mycobacterium avium subspecies paratuberculosis (MAP). Paratuberculosis is not treatable; therefore, the early identification and isolation of infected animals is a key point to reduce its incidence. In this paper, we analyse RNAseq experimental data of 5 ELISA-negative cattle exposed to MAP in a positive herd, compared to 5 negative-unexposed controls. The purpose was to find a small set of differentially expressed genes able to discriminate between exposed animals in a preclinical phase from non-exposed controls. Our results identified 10 transcripts that differentiate between ELISA-negative, clinically healthy, and exposed animals belonging to paratuberculosis-positive herds and negative-unexposed animals. Of the 10 transcripts, five (TRPV4, RIC8B, IL5RA, ERF, CDC40) showed significant differential expression between the three groups while the remaining 5 (RDM1, EPHX1, STAU1, TLE1, ASB8) did not show a significant difference in at least one of the pairwise comparisons. When tested in a larger cohort, these findings may contribute to the development of a new diagnostic test for paratuberculosis based on a gene expression signature. Such a diagnostic tool could allow early interventions to reduce the risk of the infection spreading.