Biology subjects

Tischler, N. D.

Publications and source records attributed to Tischler, N. D..

4 recordsLinked to original sources

bioRxiv2026

A historical cross-border Andes virus lineage reveals the origin of a cruise ship hantavirus pulmonary syndrome outbreak

Andes virus (ANDV) caused a multi-country outbreak of hantavirus pulmonary syndrome among passengers and crew of a cruise ship in 2026. To investigate the origin and evolutionary history of the virus responsible for the outbreak, we analyzed complete ANDV small (S), medium (M), and large (L) genome segment sequences from Chile alongside outbreak-associated and publicly available genome sequences. Across all three segment-specific phylogenies, the outbreak virus clustered within an ANDV Clade III cluster spanning southern Chile and northern Patagonia in Argentina and were most closely related to a human-derived ANDV (p1236) collected in Los Rios Region of Chile in 2012, representing the closest known historical relative of the outbreak-associated virus. Phylogeographic analysis showed that the genetically distinct ANDV Clade V lineage circulating in central Chile was not closely related to the cruise ship outbreak-associated genomes, thereby reducing the likelihood that the index cases acquired infection through zoonotic spillover while traveling through the Maule Region. These findings trace the geographic origin of the outbreak-associated virus to a defined corridor, the Hua Hum Pass, a cross-border zone connecting Province of Neuquen in Argentina with the Los Rios and La Araucania Regions of Chile.

evolutionary biology↗

bioRxiv2026

Specific determinants of the Transmembrane region of the Andes virus Gc glycoprotein drive the transition from membrane hemifusion to pore formation

Andes virus (ANDV), a highly pathogenic orthohantavirus, enters host cells through low pH-triggered membrane fusion mediated by the Gc glycoprotein, a class II fusion protein containing a single C-terminal transmembrane domain (TMD). While the ectodomain has been extensively characterized, the role of the TMD in late-stage fusion remains unclear. Here, we investigated the minimal functional length and sequence requirements of the ANDV Gc TMD using site-directed mutagenesis. C-terminal deletion mutants and serine-to-alanine substitutions were evaluated for protein expression, virus-like particle production, cell-cell fusion, pseudotyped vector entry, and hemifusion activity. Deletion of the Gc cytoplasmic tail (CT) or a single C-terminal TMD residue was tolerated, whereas deletion of two or more residues impaired particle production and fusion, indicating that at least 21 of the 22 TMD residues are required for efficient membrane fusion and viral entry. Hemifusion assays showed that deletion of two or three residues, or substitution of the strictly conserved S1121, allowed lipid mixing but blocked progression to full fusion, while deletion of four residues also abolished hemifusion. In contrast, mutation of the less conserved S1126 had minimal effect. These results identify a precise TMD length and a conserved polar TMD residue as critical determinants of fusion pore formation in ANDV.

microbiology↗

bioRxiv2025

Cell surface localization and intercellular transfer of the hantavirus nucleocapsid protein via a viral assembly-independent mechanism

Hantaviruses are segmented negative-sense RNA viruses that package their genome into helical ribonucleocapsids, by oligomerization of the nucleocapsid (N) protein along the viral RNA. Upon transmission to humans, hantaviruses can cause severe disease characterized by increased vascular permeability in the microvasculature, a key driver of pathogenesis. In patients and rodent reservoirs, robust and persistent antibody responses against the nucleocapsid (N) protein are consistently observed. Here we investigated whether the hantavirus N protein, described as a strictly intracellular protein, is released into the extracellular environment during viral infection. By using diverse experimental approaches, we demonstrate that in cells infected with Andes virus (ANDV) or the non-pathogenic Tula virus (TULV), the N protein localizes on the plasma membrane. Transfection experiments further revealed that the surface localization of N proteins from Murine, Arvicoline, Neotomine and Sigmodontine rodent-reservoir-associated hantaviruses did not require co-expression of other viral proteins indicating that its trafficking to the cell surface is mediated by a mechanism distinct from canonical virion assembly pathways. Moreover, we show that the N protein is secreted from cells into the extracellular milieu and is further transferred to neighboring cells. Interestingly, despite being present on the cell surface and secreted into the extracellular media, the TULV N protein transfer of the N protein to neighboring cells was significantly lower compared to that of ANDV N protein. The extracellular presence of hantavirus N proteins, and likely also from other clinically-relevant bunyaviruses, unveils new possible functions of the protein during the viral infection cycle, which is likely related to immune responses, immune modulation and viral pathogenesis.

microbiology↗

bioRxiv2020

Detection of Envelope Glycoprotein Assembly from Old-World Hantaviruses in the Golgi Apparatus of Living Cells

Hantaviruses are emerging pathogens that occasionally cause deadly outbreaks in the human population. While the structure of the viral envelope has been characterized with high precision, the protein-protein interactions leading to the formation of new virions in infected cells are not fully understood yet. In this work, we use quantitative fluorescence microscopy (i.e. Number&Brightness analysis and fluorescence fluctuation spectroscopy) to quantify the interactions that lead to oligomeric spike complex formation in the physiological context of living cells. To this aim, we have analyzed proteins from Puumala and Hantaan orthohantaviruses in several cellular models. For the first time, we quantified the oligomerization state of each protein in relation to its subcellular localization, concentration and the concentration of its interaction partner. Our results indicate that when expressed separately, both glycoproteins can form homo-multimers in a concentration-dependent manner. Fluorescence fluctuation analysis was applied to prove that Gc:Gc contacts observed on virions are also relevant for Gc-Gc interactions in living cells, in the absence of Gn. Furthermore, we proved that the membrane-distal lobes of Gn are not necessary for Gn homo-multimerization. In cells co-expressing both glycoproteins, we observe clear indication of Gn-Gc interactions and the formation of protein complexes with different sizes, while using various labelling schemes to minimize the influence of the fluorescent tags. Our data are compatible with an assembly model according to which hantavirus spikes are formed via the assembly of Gn-Gc hetero-dimers. Furthermore, our results indicate the interconnection of large Gn-Gc hetero-multimers in the Golgi apparatus. Such large glycoprotein multimers may be identified as multiple interacting viral spikes and provide a possible first evidence for the initial assembly steps of the viral envelope, within this organelle, directly in living cells.

biophysics↗