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Mechanism of polarization of Listeria monocytogenes surface protein ActA

Mechanism of polarization of Listeria monocytogenes surface protein ActA. MM 59 , 1262-1279. The life cycle. Nat. Rev. Microl . 4 , 423 (2006). JAT-396. JAT-395. 20-30min. 30-60min. 1-3hr. 3-6hr. I II III IV . A delay? II-IIII accumulation and III-IV redistribution?

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Mechanism of polarization of Listeria monocytogenes surface protein ActA

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  1. Mechanism of polarization of Listeriamonocytogenessurface protein ActA MM59, 1262-1279

  2. The life cycle Nat. Rev. Microl. 4, 423 (2006)

  3. JAT-396 JAT-395

  4. 20-30min 30-60min 1-3hr 3-6hr

  5. I II III IV A delay? II-IIII accumulation and III-IV redistribution? Slow kinetics of ActA rearrangement derived from “growth of cell wall”

  6. For Bacteria entry and host specificity Nat. Rev. Microl. 4, 423 (2006)

  7. Time-Course of InlA

  8. partial helical pattern Secretion of ActA-RFP occurred at sites distinct from new cell wall synthesis

  9. The percentage of poles associated with ActA-RFP was greatest for the youngest

  10. Retention of ActA at inert poles

  11. Actin-based motility Burkholderiapseudomallei Nat. Rev. Microl. 4, 91 (2006)

  12. Actin based motility and ActA distribution Black outline represents the bacterium. The polar ActA distribution on the surface is shown in blue. The actin that polymerized first (oldest actin) is shown in red, and the newest is in yellow. The dashed line indicates the stationary laboratory frame of reference. Initial actin accumulation (red) occurs preferentially along the sides as the hemispherical poles have a disadvantage toward actin accumulation due to their greater curvature. Biophysical J89, 2146 (2005)

  13. Oldest poles took the longest to support motility with interruptions 2:20:00 2:20:00 Less than 1 hour

  14. Progress Report

  15. cytology Fixed cells parA localization ΔminCDE deplete MreB FtsZ/Ats; Aztreonam Effect on nuclear Live cells oscillation pattern Recombinant protein Functional operon parSA1B in pxx326 (F-plasmid) Solubility with tags CBP GST NUS His malE Trx

  16. pLE7/MC1000 0min 10min A22 -A22

  17. pSOT27/MC1000 0min 10min A22 -A22

  18. pSOT27/ JFL, OV16 pMLB1113/MC1000 pSOT27/JFL AztreonamIPTG 40min pSOT27/OV16

  19. 10min 1hr 2hr

  20. pSOT27/MC1000 Treating with Aztreonam

  21. pSOT27/MC1000 Increase treating time IPTG 20μM (min) 100 130 150

  22. AZT IPTG 90min AZT IPTG 70min

  23. On Going • F-plasmid stability • parA recombinant protein solubility • parAlocalization in filamentous cells • Effect on nucleoid (nuclear-cell length)

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