Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2023 Sep 6;14:5475. doi: 10.1038/s41467-023-41174-0

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 4 — a Maximum-likelihood phylogenetic tree of tent genes including novel tent sequences assembled from ancient DNA samples and a non-redundant set of tent sequences from existing strains in which duplicates have been removed (see “Methods” for details). The phylogeny has been subdivided into three subgroups. Sequences are labeled according to sample followed by their associated clade in the genome-based tree (Fig. 2a), except for the Barcelona-3031-Tooth sequence (*) as it fell below the coverage threshold. b Visualization of tent sequence variation, with vertical bars representing nucleotide substitutions found uniquely in tent sequences from ancient DNA samples. On the right, a barplot is shown that indicates the number of unique substitutions found in each sequence, highlighting the uniqueness of subgroup 3. c Structural model of TeNT/Chinchorro indicating all of its unique amino acid substitutions, which are not observed in modern TeNT sequences. Also shown is a segment of the translated alignment for a specific N-terminal region of the TeNT protein (residues 141–149, Uniprot ID P04958). This sub-alignment illustrates a segment containing a high density of unique amino acid substitutions, four of which are shared in TeNT/El-Yaral and TeNT/Chinchorro. d MapDamage analysis of the tent/Chinchorro gene, and associated C. tetani contigs and mtDNA from the Chinchorro-Mummy-Bone sample. e Cultured rat cortical neurons were exposed to full-length toxins in culture medium at the indicated concentration for 12 h. Cell lysates were analyzed by immunoblot, and the image shown is a representative of four independent experiments. WT TeNT (uniprot accession # P04958) and TeNT/Chinchorro (“ch”) showed similar levels of activity in cleaving VAMP2 in neurons. f, g Full-length toxins ligated by sortase reaction were injected into the gastrocnemius muscles of the right hind limb of mice. The extent of muscle rigidity was monitored and scored for 4 days (means ± s.e.; n = 3 per group, 9 total). TeNT/Chinchorro (“ch”) induced typical spastic paralysis and showed a potency similar to WT TeNT. Source data for (a, b, d, e, g) are provided as a Source Data file.