Small Ubiquitin-related Modifier Ligase Activity of Mms21 Is Required for Maintenance of Chromosome Integrity during the Unperturbed Mitotic Cell Division Cycle in Saccharomyces cerevisiae

The SUMO ligase activity of Mms21/Nse2, a conserved member of the Smc5/6 complex, is required for resisting extrinsically induced genotoxic stress. We report that the Mms21 SUMO ligase activity is also required during the unchallenged mitotic cell cycle in Saccharomyces cerevisiae. SUMO ligase-defective cells were slow growing and spontaneously incurred DNA damage. These cells required caffeine-sensitive Mec1 kinase-dependent checkpoint signaling for survival even in the absence of extrinsically induced genotoxic stress. SUMO ligase-defective cells were sensitive to replication stress and displayed synthetic growth defects with DNA damage checkpoint-defective mutants such as mec1, rad9, and rad24. MMS21 SUMO ligase and mediator of replication checkpoint 1 gene (MRC1) were epistatic with respect to hydroxyurea-induced replication stress or methyl methanesulfonate-induced DNA damage sensitivity. Subjecting Mms21 SUMO ligase-deficient cells to transient replication stress resulted in enhancement of cell cycle progression defects such as mitotic delay and accumulation of hyperploid cells. Consistent with the spontaneous activation of the DNA damage checkpoint pathway observed in the Mms21-mediated sumoylation-deficient cells, enhanced frequency of chromosome breakage and loss was detected in these mutant cells. A mutation in the conserved cysteine 221 that is engaged in coordination of the zinc ion in Loop 2 of the Mms21 SPL-RING E3 ligase catalytic domain resulted in strong replication stress sensitivity and also conferred slow growth and Mec1 dependence to unchallenged mitotically dividing cells. Our findings establish Mms21-mediated sumoylation as a determinant of cell cycle progression and maintenance of chromosome integrity during the unperturbed mitotic cell division cycle in budding yeast.

Expediency of Penetration Ratio and Evaluation of Mean Throughput for Safety and Commercial Applications in VANETs

Vehicular ad hoc network (VANET) applications are principally categorized into safety and commercial applications. Efficient traffic management for routing an emergency vehicle is of paramount importance in safety applications of VANETs. In the first case, a typical example of a high dense urban scenario is considered to demonstrate the role of penetration ratio for achieving reduced travel time between source and destination points. The major requirement for testing these VANET applications is a realistic simulation approach which would justify the results prior to actual deployment. A Traffic Simulator coupled with a Network Simulator using a feedback loop feature is apt for realistic simulation of VANETs. Thus, in this paper, we develop the safety application using traffic control interface (TraCI), which couples SUMO (traffic simulator) and NS2 (network simulator). Likewise, the mean throughput is one of the necessary performance measures for commercial applications of VANETs. In the next case, commercial applications have been considered wherein the data is transferred amongst vehicles (V2V) and between roadside infrastructure and vehicles (I2V), for which the throughput is assessed.

Operational inferences on VANETs in 802.16e and 802.11p with improved performance by Congestion alert

Mobile WiMAX is a burgeoning network technology with diverse applications, one of them being used for VANETs. The performance metrics such as Mean Throughput and Packet Loss Ratio for the operations of VANETs adopting 802.16e are computed through simulation techniques. Next we evaluated the similar performance of VANETs employing 802.11p, also known as WAVE (Wireless Access in Vehicular Environment). The simulation model proposed is close to reality as we have generated mobility traces for both the cases using a traffic simulator (SUMO), and fed it into network simulator (NS2) based on their operations in a typical urban scenario for VANETs. In sequel, a VANET application called `Street Congestion Alert' is developed to assess the performances of these two technologies. For this application, TraCI is used for coupling SUMO and NS2 in a feedback loop to set up a realistic simulation scenario. Our inferences show that the Mobile WiMAX performs better than WAVE for larger network sizes.