7 research outputs found
Molecular signatures for CCN1, p21 and p27 in progressive mantle cell lymphoma
Mantle cell lymphoma (MCL) is a comparatively rare non-Hodgkin’s lymphoma characterised by overexpression of cyclin D1.Many patients present with or progress to advanced stage disease within 3 years. MCL is considered an incurable disease withmedian survival between 3 and 4 years. We have investigated the role(s) of CCN1 (CYR61) and cell cycle regulators inprogressive MCL. We have used the human MCL cell lines REC1 G519 > JVM2 cells by RQ-PCR, depicting a decrease in CCN1expression with disease progression. Investigation of CCN1 isoform expression by western blotting showed that whilst expres-sion of full-length CCN1 was barely altered in the cell lines, expression of truncated forms (18–20 and 28–30 kDa) decreasedwith disease progression. We have then demonstrated that cyclin D1 and cyclin dependent kinase inhibitors (p21CIP1and p27KIP1)are also involved in disease progression. Cyclin D1 was highly expressed in REC1 cells (OD: 1.0), reduced to one fifth in G519cells (OD: 0.2) and not detected by western blotting in JVM2 cells. p27KIP1followed a similar profile of expression as cyclin D1.Conversely, p21CIP1was absent in the REC1 cells and showed increasing expression in G519 and JVM2 cells. Subcellularlocalization detected p21CIP1/p27KIP1primarily within the cytoplasm and absent from the nucleus, consistent with altered roles in treatment resistance. Dysregulation of the CCN1 truncated forms are associated with MCL progression. In conjunction withreduced expression of cyclin D1 and increased expression of p21, this molecular signature may depict aggressive disease andtreatment resistance
Covalent Inhibition of Ubc13 Affects Ubiquitin Signaling and Reveals Active Site Elements Important for Targeting
Ubc13 is an E2 ubiquitin
conjugating enzyme that functions in nuclear
DNA damage signaling and cytoplasmic NF-κB signaling. Here,
we present the structures of complexes of Ubc13 with two inhibitors,
NSC697923 and BAY 11-7082, which inhibit DNA damage and NF-κB
signaling in human cells. NSC697923 and BAY 11-7082 both inhibit Ubc13
by covalent adduct formation through a Michael addition at the Ubc13
active site cysteine. The resulting adducts of both compounds exploit
a binding groove unique to Ubc13. We developed a Ubc13 mutant which
resists NSC697923 inhibition and, using this mutant, we show that
the inhibition of cellular DNA damage and NF-κB signaling by
NSC697923 is largely due to specific Ubc13 inhibition. We propose
that unique structural features near the Ubc13 active site could provide
a basis for the rational development and design of specific Ubc13
inhibitors