The Terminal List (S01) __EXCLUSIVE__
On paper, the premise is simple and effective, with the idea of a kill list essentially working as an ever-evolving mission for Reece to conduct. As the episodes tick by, and more and more names are added and crossed off that list, The Terminal List begins to slow down and become a laborious chore.
The Terminal List (S01)
Reece is drawn deeper into a complex conspiracy and puts... More a new name on his list. Meanwhile, Secretary of Defense Lorraine Hartley announces a drastic policy overhaul to help Special Operators. And Katie discovers the truth of what's going on in Reece's head.
The Terminal List (ou La Liste terminale au Québec) est une série télévisée américaine basée sur le roman du même nom écrit par Jack Carr. Développée par David DiGillo, elle est diffusée pour la première fois dans le monde entier sur Prime Video à partir du 1er juillet 2022.
OG18056 is an active NADase that produces a variant of cADPR. (A) Alignment of a structural model of OG18056 (predicted by AlphaFold2, in cyan) with a human ADPR cyclase CD38 (6VUA, in gray) and the TIR domain of human SARM1 (6O0R, 561 to 700 aa, in light brown). For visualization purposes, sequences outside the alignment region were removed, and those that did not align within the region are more transparent. The position of the predicted catalytic Glu residues, which are aligned in the three proteins, is highlighted with a box. (B) A close-up of alignments of OG18056 with TIR-hSARM1 (Upper) and hCD38 (Lower) at the predicted catalytic site showing a Glu E305 in OG18056 that is aligned with the catalytic residues E642 in TIR-hSARM1 and E226 in hCD38. (C) Electrostatic image of the surface of the structural model of OG18056 with regions of positive, neutral, and negative charge colored in blue, white, and red, respectively. A molecule of NAD+ was docked into the predicted catalytic pocket. (D) HPLC chromatograms showing the cADPR product produced by OG18056 using a semi-in vitro assay. OG18056 and OG18056E305A were expressed in N. benthamiana. After immunoprecipitation, the proteins were incubated with NAD+ for 1 h before subjection to LCMS analysis. The controls were NAD+ only without addition of beads and immunoprecipitation (IP) of proteins from leaves infiltrated with Agrobacterium carrying an EV. Peaks corresponding to m/z 542.06 (cADPR) and m/z 664.11 (NAD+) were identified. (E) Immunoblots detecting the expression of OG18056 and OG18056E305A in N. benthamiana (input) and after IP. The IP samples were then used for the LCMS analyses. OG18056 and OG18056E305A were tagged with C-terminal 3FLAG. Leaves infiltrated with Agrobacterium carrying the empty vector (EV) was included as a control. Ponceau Staining (PS) of the membrane was used as a loading control.
Using BLASTP, we identified homologs of HopBY in diverse bacterial lineages ranging in amino acid sequence identity from 26 to 100%. HHsearch confirmed that these homologs all have an ADPR cyclase-like fold on their C terminus (Dataset S3). Examination of the genomic region surrounding all homologs revealed they are often associated with transposons, indicating a strong signature of horizontal gene transfer (SI Appendix, Fig. S10B). A phylogeny based on structure-guided protein sequence alignment grouped these HopBY homologs into two clades (Fig. 5A). The HopBY from Pseudomonas syringae is grouped with homologs from other Pseudomonas spp. as well as plant pathogenic Enterobacteriaceae species within the genera Brenneria and Pectobacterium. All these genomes contain genes encoding a Hrp1 T3S, the canonical T3S found in Pseudomonas syringae. Importantly, the homologous genes in this clade are all preceded by a hrp box promoter (Dataset S3), indicating that they are likely T3Es. In the Brenneria (n = 2) genomes, the hopBY homologous genes are present close to the T3S machinery gene clusters, further supporting their identity as T3Es. The other clade includes more distinct HopBY homologs including one in Paraburkholderia sp. ZP32-5, which was also predicted as a possible T3E. Paraburkholderia sp. ZP32-5 contains the Hrp2 type of T3S machinery and the hopBY homolog gene is located in close proximity to the T3S gene cluster. Upstream of this gene, there are PIP-box motifs, which are associated with Hrp2 T3S-regulated gene expression. Therefore, this HopBY homolog is likely also a T3E. Interestingly, other homologs in this clade are often located adjacent to type VI secretion system (T6S) genes such as vgrG, tssA, vasA, and tssG. T6S is an important secretion system for bacterial interactions with other bacteria and eukaryotes by delivering toxins that kill recipient cells (39). Indeed, these more distant homologs of the Pseudomonas syringae HopBY were predicted to be putative T6S effectors. Some of these proteins have domains such as the N-terminal Rhs (Rearrangement Hotspot) core domain, which are often found in T6Es (40). 041b061a72