Persister Cells Resuscitate Using Membrane Sensors that Activate Chemotaxis, Lower cAMP Levels, and Revive Ribosomes

Abstract

Persistence, the stress-tolerant state, is arguably the most vital phenotype since nearly all cells experience nutrient stress, which causes a sub-population to become dormant. However, how persister cells wake to reconstitute infections is not understood well. Here, using single-cell observations, we determined that Escherichia coli persister cells resuscitate primarily when presented with specific carbon sources, rather than spontaneously. In addition, we found that the mechanism of persister cell waking is through sensing nutrients by chemotaxis and phosphotransferase membrane proteins. Furthermore, nutrient transport reduces the level of secondary messenger cAMP through enzyme IIA; this reduction in cAMP levels leads to ribosome resuscitation and rescue. Resuscitating cells also immediately commence chemotaxis toward nutrients, although flagellar motion is not required for waking. Hence, persister cells wake by perceiving nutrients via membrane receptors that relay the signal to ribosomes via the secondary messenger cAMP, and persisters wake and utilize chemotaxis to acquire nutrients.

Combatting Persister Cells With Substituted Indoles

“…This heterogeneous nature of persister cell resuscitation was subsequently verified by others (Goormaghtigh and Van Melderen, 2019;Pu et al, 2019). Using single cells and searches over all E. coli proteins, we determined that persister cell resuscitation is initiated by recognizing external nutrients through receptors for chemotaxis (for amino acids) and phosphotransferase membrane proteins (for glucose) and does not require proteins specialized for persistence ( Figure 1B) (Yamasaki et al, 2020). Resuscitation is also not primarily spontaneous but instead is based on the recognition of nutrients (Yamasaki et al, 2020).…”

“…Using single cells and searches over all E. coli proteins, we determined that persister cell resuscitation is initiated by recognizing external nutrients through receptors for chemotaxis (for amino acids) and phosphotransferase membrane proteins (for glucose) and does not require proteins specialized for persistence ( Figure 1B) (Yamasaki et al, 2020). Resuscitation is also not primarily spontaneous but instead is based on the recognition of nutrients (Yamasaki et al, 2020). The presence of external nutrients (i.e., signals) is propagated to the cytosol by reducing concentrations of the secondary messenger cAMP; reduction in cAMP allows ribosomes stalled on mRNA to be rescued and inactive 100S ribosomes to be activated by HflX ( Figure 1B) (Yamasaki et al, 2020).…”

Reactive Oxygen Species Penetrate Persister Cell Membranes of Escherichia coli for Effective Cell Killing

“…However, no studies have evaluated the sterilization of persister cells using ROS externally. To determine whether ROS are effective against persister cells, we prepared rifampicininduced E. coli persister cells as previously described (Kwan et al, 2013;Kim et al, 2018a,b;Yamasaki et al, 2020). The RVR can be used in various modes such as UV mode and O 2 plasma mode.…”

Copper Kills Escherichia coli Persister Cells

“…Realizing genetic resistance could limit the use of copper as an effective antimicrobial, we assessed the likelihood of copper resistance using pooled libraries [12] of the 3985 KEIO E. coli single-gene knockouts which contains all non-lethal mutations of E. coli K-12 [31] as well as pooled libraries [2] of the ASKA plasmid library [53] where all of the 4287 E. coli proteins are produced. We reasoned that either a deletion or production of a key protein would allow higher copper tolerance and that this approach could also give us additional clues about the mode of action of copper.…”

“…Persister cells are a subpopulation of phenotypic variants that are tolerant to antimicrobial compounds. This phenotype was discovered in the early 1940s [1] and occurs primarily as a stress response [2][3][4]. Since they are metabolically inactive [5,6], antibiotics often do not eradicate these cells.…”