CACAO Phage Hunters 2018
My Annotations
| Status | Page | Date/Time | GO Term (Aspect) | Reference | Evidence | Notes | Links |
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| acceptable | LAMBD:ENLYS | 2018-03-04 23:17:00 CST | GO:0044659 cytolysis by virus of host cell (P) | PMID:28637001 | ECO:0000314 direct assay evidence used in manual assertion | The R lysis cassette, including R endolysin and S holin genes was synthesized and inserted into a plasmid under regulation of arabinose and temperature sensitive promoters. The plasmid was introduced into Δasd E. coli χ6212 and the construct was named JOL1675. Under conditions inducing expression of the plasmid, presence of the R endolysin was confirmed by Western Blot and shown in Figure 2. A band at 17.7 kDa, the estimated size of the endolysin, was seen in cells with the plasmid expressed, but not in cells grown under conditions repressing expression. Lysis efficiency was confirmed by monitoring the number of viable cells, which declined to 0 after 48 hours of lysis, as shown in Figure 4. No viable cells remained, indicating that the endolysin effectively ruptured and killed the hosts.
| challenge |
| acceptable | LAMBD:HOLIN | 2018-03-14 09:03:56 CDT | GO:0045918 negative regulation of cytolysis (P) | PMID:21187415 | ECO:0000314 direct assay evidence used in manual assertion | Figure 3. S105, holin, and S107, antiholin, are two transmembrane proteins produced by alternate starts on gene S in phage lambda. S105 accumulates into rafts, which are associated with the formation of large holes in the membrane. These holes allow endolysin to reach the peptidoglycan and result in cytolysis. S105 has three transmembrane domains. The difference between S105 and S107 is an extra N-terminal positive charge on S107, which prevents the first transmembrane domain from entering the membrane. The researchers created a mutant allele of S105 with the first transmembrane domain deleted, such that it would act like S107, and fused it to mCherryFP. Through deconvolution fluorescence imaging, they showed that the fusion protein blocked S105 raft formation, suggesting that antiholins prevent lysis by blocking the formation of holin rafts.
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unacceptable:0
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Annotations challenged by CMattson
| Status | Author,Group | Page | GO Term (Aspect) | Reference | Evidence | Links | Page history |
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