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TechSpark is a national civic program aimed at fostering greater economic opportunity and job creation through partnership with rural and smaller metropolitan communities. Is also the founder of Proper Printshop Productions and previously worked as a Financial Advisor for Per Stirling and Raymond James. He is actively involved in the community and serves on the board of directors for Fab Lab El Paso, The Borderplex Alliance Entrepreneurship Taskforce, and City of El Paso Regional Renewable Energy Advisory Council.

This experiment was performed three times¤ cutting out different numbers of gel bands, and proteins that were seen more than once in LC/MS runs are noted with a superscript r . For functional assembly reactions, we added Alexa 594-tubulin to visualize microtubule assembly in concentrated lysate, incubated reactions in tubes, and spotted aliquots onto slides for visualization. HeLa S3 cells were treated with 10 μM S-trityl-L-cysteine for 11 h and then 1 μg/ml latrunculin A for 1 h at 37°C before generation of lysates. Cells were pelleted at 400 × g for 5 min and washed three times in ice-cold HBS. Pellets were flash-frozen in liquid nitrogen and thawed at 30°C twice and then centrifuged at 5000 × g at 4°C for 30 min. Supernatants were obtained and stored on ice, and 1 μg/ml cytochalasin B was added.

Poly(ADP-ribose) , made by PARP-5a/tankyrase-1, localizes to the poles of mitotic spindles and is required for bipolar spindle assembly, but its molecular function in the spindle is poorly understood. To investigate this, we localized pADPr at spindle poles by immuno-EM. We then developed a concentrated mitotic lysate system from HeLa cells to probe spindle pole assembly in vitro.

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As a control, we expressed a GFP fusion of catalytically inactive PARP-5a (GFP-PARP-5a PD) and purified it from lysate on magnetic beads using the same procedure. These beads are coated with PARP-5a polypeptide, but not with pADPr chains. Beads coated with catalytically inactive PARP-5a were unable to assemble microtubule asters in concentrated mitotic lysate . Beads coated with mitotic PARP-5a from which pADPr chains were extended induced robust formation of microtubule asters , suggesting that the presence of pADPr on PARP-5a is required for microtubule aster assembly around the beads.

• Throughout our extensive career within the diversified and ever-changing construction/maintenance industry, we continue to maintain a competitive edge in providing services and products that utilizing the latest technology.
• If the primary function of PARP-5a is to ensure that spindle assemble only two poles, one might expect its activity to be up-regulated during mitosis.
• On obtaining a positive result, we went on to identify noncovalent pADPr-binding proteins in the lysate that might mediate spindle pole organization.
• In all cases examined, the human cell extracts behaved similar to Xenopus laevis egg extract spindle pole assembly reactions.
• We performed several LC/MS experiments, and most of the proteins in Table 1 were observed in more than one experiment, as indicated with a superscript r in Table 1.

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Her career began at Rudolph Miles and Sons, a custom house brokerage company, and has culminated at RMPersonnel, a staffing company that she has successfully managed since its inception in 1990. Ceci spends numerous hours volunteering in many community and non-profit organizations. Currently, she serves on the board of directors for the Borderplex Bi-National Economic Alliance, the Burkitt Foundation and various other advisory boards. She is a director emeritus of the El Paso Zoological Society and was honored in 2006 by UTEP’s College of Education with the Gold Nugget Award and as a Distinguished Alumni in 2010. John Mack oversees global business & technology solutions in Ireland and El Paso for Prudential. He is also on the company’s Corporate Social Responsibility & Community Engagement Council. Prior to this role, he worked as a IT Manager for Prudential, Senior Project Manager for Fannie Mae, IT Project Manager for GECU, and Operations Specialist for Bank of America.

It is important to note that all known pADPr antibodies fail to detect polymers shorter than ∼10 ADPr residues. Given that the PARP-5a used in our assay is purified from mitotic extract, it is probable that a relevant amount if pADPr is associated with PARP-5a in the 0 μM NAD+, but is not visualized on the pADPr immunoblot.

El Paso’s growth in recent years has been driven largely by the major expansion of Fort Bliss, which is widely considered to be the most valuable military installation in North America. The Borderplex Alliance, a regional economic development group, is also working aggressively to attract other businesses in a variety of sectors, including aerospace and defense, life sciences, advanced manufacturing and logistics, business services, and tourism.

West is also committed to providing a drug-free workplace and works to enforce and ensure reduction of harm to self and others. G.A. West prides itself in being a user-friendly contractor committed to transparency, responsiveness and flexibility to meet the ever-changing needs of our clients. We work to provide the highest degree of safety, quality and ethical behavior. Our commitment https://adprun.net/ to safety and safe employees is very important to us; we work to ensure no harm to any individual or the environment. ADP® Offers Important Year-End Payroll and Tax Tips to Small Business Owners and Their Advisors accountant or payroll provider of the updated information. Please visit ADP’s accountant portal (adp.com/accountant), which includes many accounting resources and tools.

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This suggests a relatively high-affinity interaction between NuMA and pADPr that is independent of the exact chain length or local density of pADPr. One of these binding proteins is NuMA, which appears to bind directly to the polymer through its rod domain, as assayed with bacteria-expressed NuMA fragments. Of these proteins, the most obvious candidate for a role in spindle pole assembly was NuMA. This protein was found in all LC/MS analyses of pADPr-binding proteins and was confirmed in multiple Western blot experiments (e.g., see Figure 5). NuMA was not found in the RNA-binding proteins from mitotic lysate as assayed by Western blot , so it binds pADPr specifically.

Prior to this role Mitzabeth worked for ADP as the Vice President for Client Services, Vice President for Migration Services, and Senior Implementation Manager. ADP is a human resources management software company that employees over 1,700 people in El Paso, including 350 UTEP graduates. ADP has given over $2.5M to UTEP all time, primarily to support the ADP Math and Science Teachers Academy. ADP provides software for the UTEP College of Business Administration and ADP Human Capital Management Academy as an in-kind donation. Although hydrolysis of pADPr is usually mediated by PARG, our finding ARH3 as a pADPr-binding protein in mitotic lysate suggest this enzyme may also participate. Omission of NAD+ and PARG inhibitor resulted in loss of most binding proteins, confirming that most of the binding proteins are recruited to pADPr on the beads, not to the PARP-5a polypeptide or the bead matrix. Mitotic lysate proteins that bound to pADPr are shown in Figure 4b, lane 3.

We use cookies to help provide and enhance our service and tailor content and ads. •PARylated CTCF changes DNA binding and localisation in a model cell line, 226LDM. After graduating from UTEP, Carmen Perez attended Southern Methodist University School of Law. Before joining Linebarger Goggan in 1995, her legal career included serving as an attorney to the Texas court of appeals, the Consumer Protection Division of the Texas Attorney General’s Office, and the El Paso County Attorney’s Office. Her community service includes the State Bar of Texas , American Bar Association, the Children’s Rights Litigation Working Group, College of the State Bar of Texas, Texas Center for Legal Ethics, and Hispanic Issues Section of the State Bar of Texas. As a member of the Mexican American Bar Association , she was honored in 2016 she with their „Investing in Our Community’s Future” award. Carmen is married to Jim Brod. She has a daughter, Jacqueline, and a son, Juan Jose.

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John is active within the community and serves on the Board of Directors for the UTEP Alumni Association and United Way of El Paso Young Leaders Society. He completed his BBA degree in Finance and Computer Information Systems at UTEP. Wilde A., Zheng Y. Stimulation of microtubule aster formation and spindle assembly by the small GTPase Ran. Wilde A., Lizarraga S. B., Zhang L., Wiese C., Gliksman N. R., Walczak C. E., Zheng Y. Ran stimulates spindle assembly by altering microtubule dynamics and the balance of motor activities. Nachury M. V., Maresca T. J., Salmon W. C., Waterman-Storer C. M., Heald R., Weis K. Importin beta is a mitotic target of the small GTPase Ran in spindle assembly. Merdes A., Ramyar K., Vechio J. D., Cleveland D. W. A complex of NuMA and cytoplasmic dynein is essential for mitotic spindle assembly. If the primary function of PARP-5a is to ensure that spindle assemble only two poles, one might expect its activity to be up-regulated during mitosis.

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In most experiments 1 mM NAD+ and 1 μM ADP-HPD were also added to protect and grow pADPr chains. If incubated with ARH3, samples were subsequently washed three times 10 min with cell lysis buffer containing 300 mM NaCl followed by one time for 10 min with cell lysis buffer and then eluted with SDS-PAGE sample buffer. To identify proteins that bind to pADPr in mitotic lysate, we performed LC/MS on gel bands excised from the experiment shown in Figure 4e, lane 4, as indicated by the boxes on the gel lane . We avoided bands that seemed to be common to the RNA and pADPr lanes and also with an abundant high-molecular-weight band that we already knew to be mostly filamin and spectrin.

Rodrigo is a member of the El Paso del Norte Group and serves on the boards of various companies in the banking and telecommunications industry. In 2001, Rodrigo graduated from UTEP with a Bachelor of Business Administration degree in finance. An engineering, business, and management consulting expert with over twenty-five years’ experience, Peter Chan started his career in Texas. His experience also includes managing large projects and starting up new business lines in water, wastewater and membrane projects for municipal and industrial clients. Peter now runs his own real estate investment firm in Arizona, where he currently manages real estate investment properties, property development, investment funding, and water and wastewater consulting. Peter enjoys staying active in the community, having established an endowed scholarship at UTEP, while serving as a guest speaker globally for engineering firms, corporations, and student groups.

El Paso was home to El Paso Corporation formerly known as El Paso Natural Gas Company. Bound proteins were eluted with hot sample buffer and resolved by SDS-PAGE. We tested the effect of varying factors such ionic strength of lysis and wash buffers, incubation time/temperature, extent of pADPr modification, and g-force during lysate clarification, so as to maximize the efficiency and specificity of the capture reaction. In Figure 4 we report data from our current protocol, acknowledging that further experimentation is required to define the affinity and specificity of candidate pADPr-binding proteins . Magnetic beads coated with PARP-5a lacking catalytic activity (PD-PARP-5a) or PARP-5a decorated with pADPr (PARP-5a-pADPr) were added to concentrated mitotic lysate containing Alexa 594 tubulin and 1 μM ADP-ADHP to inhibit PARG activity. Note that pADPr-coated beads induced assembly of microtubule asters. Addition of extra NAD+ (+250 μM NAD+ and + 500 μM NAD+) to the lysate to promote additional pADPr polymerization increased the density of asters and the density of microtubules within them.

We use cookies to improve security, personalize the user experience, enhance our marketing activities and for other business use. The Medical Center of the Americas Foundation, a 5013 nonprofit organization, is working to lead the development of the MCA campus and advance biomedical innovation in the Paso del Norte region. The Foundation’s mission is to position the Paso del Norte region as the global leader of health delivery, education and research, concentrating on issues unique to the Hispanic, border and military populations. MCA development is expected to attract many healthcare-related research and development businesses to the area.

Asters assembled around the PARP-5a-pADPr beads were always located close to beads or bead aggregates, but the microtubules did not seem to emanate directly from the beads. Rather, the asters had distinct foci and appeared to associate laterally with the beads near these foci. To determine if the amount of pADPr on the beads affects microtubule aster assembly, we added increasing concentrations of NAD+ to the reaction . Addition of 250 μM and 500 μM NAD+ resulted in a concentration-dependent increase in the number of microtubule asters found associated with the beads, and the microtubule density in each aster . These data show that pADPr elongated from PARP-5a is capable of inducing assembly of pole-like microtubule asters in mitotic cytoplasm, arguing that it has a functional role in pole assembly in cells. We hypothesized that induction of microtubule asters by pADPr-coated beads depended on recruiting specific proteins from the mitotic lysate onto the pADPr polymers and set out to identify these proteins, as describe below. These approximately correspond to the domain organization of NuMA, representing the C- and N-terminal globular domains, and to a central, coiled-coil rod domain.

More than 70 Fortune 500 companies have offices in El Paso, including Western Refining, AT&T, ADP, Helen of Troy, Boeing, Delphi, Dish Network, Eureka, Hoover, Raytheon, State Farm and USAA. Police restored access to a major oil depot near Marseille after workers blocked it on Thursday, and the authorities also removed blockades at a depot near the south-western city of Bordeaux and at a third site near Clermont-Ferrand. Power up your marketing and get people to pay attention to your business, pursuit, or clients. Find prospects, develop your lists, and track your marketing campaigns without even having to leave the RocketReach suite. Find the most crucial people you need to bring your product to with our advanced search features and then immediately take action, leaving your competition in the dust. If you are a first time user you will need to register your account or create a new one, otherwise you can login on Andeavor Page with your previous login details. By using this site, you are agreeing to security monitoring and auditing.

Microtubule appeared not to emanate directly from the pADPr-coated beads; rather they assembled into asters that associated laterally with beads. To test if pADPr chains growing from PARP-5a can function in pole organization, we developed a functional assay in mitotic lysate. On obtaining a positive result, we went on normal balance to identify noncovalent pADPr-binding proteins in the lysate that might mediate spindle pole organization. One of these was NuMA, which thus appears to interact both covalently and noncovalently with pADPr to promote spindle pole organization. Recruitment of NuMA and CH-TOG to pADPr beads at different polymer levels.

All of these are abundant and are common contaminants in immunopurification assays. This might reflect nonspecific binding or possibly an RNA-mimic function of pADPr.

Author: Barbara Weltman