Category:Team Mom Genes

Figure 4 shows Rsp4 mutants have impaired retrograde trafficking from Golgi to ER. Figure 7 shows similar evidence.

Figure 1 shows that Sla1 and Las17 arrive and disperse simultaneously during endocytosis and form a stable complex in the cytosol; this was observed using microscopy. Figure 3 shows use of a GST fusion affinity assay to demonstrate interaction between Sla1 and Las17 is "direct and strong".

Figure 2 gives evidence of Lsb1 and Lsb2 inhibiting Las17. Figure 4 shows Lsb1 and Lsb2 are colocalized with Las17, a protein involved in actin polymerization.

Figure 8 shows overproduction of inositol in an Opi1 mutant.

Figure 5: when expressed from plasmids, dkgA was found to decrease furfural tolerance. Figure 6: Deletions of dkgA caused an increase in furfural tolerance and a decrease in furfural reductase activity in vivo similar to the results shown in the EMFR9 strain of E. coli.

Since deletion of dkgA alone did not lower the in vivo reductase activity or increase furfural tolerance, another gene, yqhD, is presumed to be the more important activity for growth inhibition by low concentrations of furfural. The lowest furfural reductase activity, however, was found after deletion of both genes as seen in Figure 3.

Fig 5 shows the results of a luciferase reactivation assay in which Clp/Hsp100 proteins can prevent the permanent inactivation of luciferase if added prior to heating. When MXAN_4832 was added before the heat shock and KJE is added after the heat shock some activity was restored. No activity was restored to luciferase when MXAN_4832 was absent before the heat shock as well as when KJE was absent after heat shock.

Fig 3A. When the agglutination rate of WT DK1622 was compared to that of Δ4832, the two rates matched one another for the first hour. After 3 hours, the agglutination rate of Δ4832 was reduced compared to that of DK1622. A reduced rate of decrease in agglutination is indicative of a defect in fibril production.

Figure 2 shows that a mutant with a deletion of MXAN_4832 exhibits a similar cell motility phenotype to that of DK1253, an A+S- strain. MXAN_4832 contributes to the regulation of S-motility because deletion of this gene causes the loss of S-motility.

Figure 2 shows DHQ formation by PqsD in Vitro. Mutant strains showed PqsD was a part of DHQ synthesis in vivo and was further tested in vitro. Anthraniloyl-CoA was produced in situ by PqsA and reacted with malonyl-CoA (a product of PqsD) to form DHQ as supported by TLC and LC-MS. These tests confirm DHQ formation when compared against the DHQ standard. Figure 7 shows the proposed mechanism for DHQ synthesis with the products of the Pqs gene.

Encodes for essential proteins involved in intracellular hopanoid transport from the cytoplasmic to the outer membrane. Figure 4 shows the content of lipid extract from membrane fractions from R. palustris compared to the lipid extract from membranes of the mutant strain (Rpal_4254) where the gene was deleted. The WT contained 12.5 +/- 1 ug x mg total lipid extract (TLE); the mutant strain (Rpal_4254) no longer contained hopanoids in the outer membrane. Both WT and Rpal_4254 showed hopanoid content in the whole cell.

Fig. 4B illustrates reduced swarm diameter observed in a frzE mutant compared to the swarm diameter observed in WT DK1622. Deletion of this gene does not completely knock out swarming ability but greatly decreases it.

Fig. 3 shows that the strain with mutations in both phosphorylation sites in frzZ exhibits the same phenotype as strains that have a complete deletion of frzZ. As seen in Fig 3B, these two strains have inhibited motility, in comparison to the wild-type strain, as evident by a smaller colony diameter on both 0.3% agar and 1.5% agar. Further, strains with a mutation in one of the possible phosphorylation sites exhibited motility similar to wild-type indicating that phosphorylation can occur at one site or the other as well as the role of frzZ in cell motility.

Fig 4B shows that radiolabelled phosphate is transferred from FrzE when in the presence of FrzZ as seen by the appearnace of a radiolabelled band in lane 3 at the FrzZ position. Fig 4B also shows that both domains of FrzZ are able to be phosphorylated by FrzE through the individual mutation of each domain at an aspartate residue, as seen by a decrease in band intensity at the FrzECheA position and the appearance of a radiolabbelled band at the FrzZ position in lanes 4 & 5. Phosphotransfer can not occur when both domains are mutated concurrently at an aspartate residue as evidenced from a lack of a radiolabelled band at the FrzZ position in lane 6.