[en] The mechanism of DNA replication is highly conserved in eukaryotes, with the process being preceded by the ordered assembly of pre-replication complexes (pre-RCs). Pre-RC formation is triggered by the association of the origin replication complex (ORC) with chromatin. Leishmania major appears to have only one ORC ortholog, ORC1. ORC1 in other eukaryotes is the largest of the ORC subunits and is believed to play a significant role in modulating replication initiation. Here we report for the first time, the cloning of ORC1 from L. major, and the analysis of its expression in L. major promastigotes. In human cells ORC1 levels have been found to be upregulated in G1 and subsequently degraded, thus playing a role in controlling replication initiation. We examine the subcellular localization of L. major ORC1 in relation to the different stages of the cell cycle. Our results show that, unlike what is widely believed to be the case with ORC1 in human cells, ORC1 in L. major is nuclear at all stages of the cell cycle

No abstract available

[en] Short communication. 5 refs, 2 figs

[en] Two key features of myeloma cells are the deregulation of the cell cycle and the dependency on the expression of the BCL2 family of anti-apoptotic proteins. The cell division cycle 7 (CDC7) is an essential S-phase kinase and emerging CDC7 inhibitors are effective in a variety of preclinical cancer models. These compounds also inhibit CDK9 which is relevant for MCL-1 expression. The activity and mechanism of action of the dual CDC7/CDK9 inhibitor PHA-767491 was assessed in a panel of multiple myeloma cell lines, in primary samples from patients, in the presence of stromal cells and in combination with drugs used in current chemotherapeutic regimens. We report that in all conditions myeloma cells undergo cell death upon PHA-767491 treatment and we report an overall additive effect with melphalan, bortezomib and doxorubicin, thus supporting further assessment of targeting CDC7 and CDK9 in multiple myeloma

[en] HPV16 E6 deregulates G1/S cell cycle progression through p53 degradation preventing transcription of the CDK inhibitor p21^WAF1. However, additional mechanisms independent of p53 inactivation appear to exist. Here, we report that HPV16 E6 targets the cellular factor p150^Sal2, which positively regulates p21^WAF1 transcription. HPV16 E6 associates with p150^Sal2, inducing its functional inhibition by preventing its binding to cis elements on the p21^WAF1 promoter. A HPV16 E6 mutant, L110Q, which was unable to bind p150^Sal2, did not affect the ability of the cellular protein to bind p21^WAF1 promoter, underlining the linkage between these events. These data describe a novel mechanism by which HPV16 E6 induces cell cycle deregulation with a p53-independent pathway. The viral oncoprotein targets p150^Sal2, a positive transcription regulator of p21^WAF1 gene, preventing G1/S arrest and allowing cellular proliferation and efficient viral DNA replication.

[en] Highlights: • Forced entry into M-phase drives replisome disassembly at stalled replication forks. • CDK activity and K48- and K63-linked ubiquitylation is required for mitotic replisome disassembly. • Mitotic replisome disassembly is mechanistically distinct from that of replication termination. The disassembly of eukaryotic replisome during replication termination is mediated by CRL-dependent poly-ubiquitylation of Mcm7 and p97 segregase. The replisome also disassembles at stalled or collapsed replication forks under certain stress conditions, but the underlying mechanism is poorly understood. Here, we discovered a novel pathway driving stepwise disassembly of the replisome at stalled replication forks after forced entry into M-phase using Xenopus egg extracts. This pathway was dependent on M-CDK activity and K48- and K63-linked poly-ubiquitylation but not on CRL and p97, which is different from known pathways. Furthermore, this pathway could not disassemble converged replisomes whose Mcm7 subunit had been poly-ubiquitylated without p97. These results suggest that there is a distinctive pathway for replisome disassembly when stalled replication forks persist into M-phase.

[en] It is currently unclear whether a correlation exists between N-myc downstream-regulated gene 2 (NDRG2) expression and oesophageal squamous cell carcinoma (ESCC). The aim of this study was to examine the underlying clinical significance of NDRG2 expression in ESCC patients and to investigate the effects of NDRG2 up-regulation on ESCC cell growth in vitro and in vivo. Immunohistochemistry was used to determine the level of NDRG2 expressions in ESCC tissue, which was then compared to specific clinicopathological features in the patient and tissue specimens. Factors associated with patient survival were analysed. Moreover, the effects of up-regulating NDRG2 expression on the growth of an ESCC cell line were examined by MTT, colony formation, DNA replication activity and nude mouse model assays. Notably low expression of NDRG2 in ESCC patients was inversely associated with clinical stage, NM classification, histological differentiation and patients’ vital status (all P < 0.05). ESCC patients expressing high levels of NDRG2 exhibited a substantially higher 5-year overall survival rate than NDRG2-negative patients. Furthermore, NDRG2 over-expression reduced the proliferation, colony formation and DNA replication activity in ESCC cells, as well as inhibiting the growth of ESCC cells in vivo. The present experiments demonstrated that NDRG2 may be a diagnostic and prognostic marker in patients with ESCC, and up-regulation of NDRG2 might act as a promising therapeutic strategy for aggressive ESCC

No abstract available

[en] How cells accomplish cell size homeostasis is a fascinating topic, and several cell size regulation mechanisms were proposed: timer, sizer, and adder. Recently the adder model has received a great deal of attention. Adder property was also found in the DNA replication cycle. This paper aims to explain the adder phenomenon both in the division-centric picture and replication-centric picture at the molecular level. We established a self-replication model, and the system reached a steady state quickly based on evolution rules. We collected tens of thousands of cells in the same trajectory and calculated the Pearson correlation coefficient between biological variables to decide which regulatory mechanism was adopted by cells. Our simulation results confirmed the double-adder mechanism. Chromosome replication initiation and cell division control are independent and regulated by respective proteins. Cell size homeostasis originates from division control and has nothing to do with replication initiation control. At a slow growth rate, the deviation from adder toward sizer comes from a significant division protein degradation rate when division protein is auto-inhibited. Our results indicated the two necessary conditions in the double-adder mechanism: one is balanced biosynthesis, and the other is that there is a protein trigger threshold to inspire DNA replication initiation and cell division. Our results give insight to the regulatory mechanism of cell size and instructive to synthetic biology. (paper)

No abstract available