Young, R. T. | bioRxiv Science

bioRxiv2025

The anaerobic gut fungal community in ostriches (Struthio camelus)

Anaerobic gut fungi (AGF; Neocallimastigomycota) are essential for plant biomass degradation in herbivores. While extensively studied in mammals, information regarding their occurrence, diversity, and community structure in non-mammalian hosts remains sparse. Here, we report on the AGF community in ostriches (Struthio camelus), herbivorous, flightless, hindgut fermenting members of the class Aves (birds). Culture-independent diversity surveys of fecal samples targeting the D2 region of the large ribosomal subunit (28S rRNA) revealed a uniform community with low alpha diversity. The community was mostly comprised of sequences potentially representing two novel species in the genus Piromyces, and a novel genus in the Neocallimastigomycota. Sequences affiliated with these novel taxa were absent or extremely rare in datasets derived from mammalian and tortoise samples, indicating a strong pattern of AGF-host association. One Piromyces strain (strain Ost1) was successfully isolated. Transcriptomics-enabled molecular dating analysis suggested a divergence time of {approx} 30 Mya, a time frame in line with current estimates for ostrich evolution. Comparative gene content analysis between strain Ost1 and other Piromyces species from mammalian sources revealed a high degree of similarity. Our findings expand the range of AGF animal hosts to include members of the birds (class Aves), highlight a unique AGF community adapted to the ostrich alimentary tract, and demonstrate that - like mammals - coevolutionary phylosymbiosis (i.e. concurrent evolution of AGF and their animal hosts) plays a central role in explaining current AGF distribution patterns in Aves.

microbiology↗

bioRxiv2021

Designed architectural proteins that tune DNA looping in bacteria

Architectural proteins alter the shape of DNA, often by distorting the double helix and introducing sharp kinks that relieve strain in tightly-bent DNA structures. Here we design and test artificial architectural proteins based on a sequence-specific Transcription Activator-like Effector (TALE) protein, either alone or fused to a eukaryotic high mobility group B (HMGB) DNA-bending domain. We hypothesized that TALE protein binding would stiffen DNA to bending and twisting, acting as an architectural protein that antagonizes the formation of small DNA loops. In contrast, fusion to an HMGB domain was hypothesized to generate a targeted DNA-bending architectural protein that facilitates DNA looping. We provide evidence from E. coli Lac repressor gene regulatory loops supporting these hypotheses in living bacteria. Both data fitting to a thermodynamic DNA looping model and sophisticated molecular modeling support the interpretation of these results. We find that TALE protein binding inhibits looping by stiffening DNA to bending and twisting, while the Nhp6A domain enhances looping by bending DNA without introducing twisting flexibility. Our work illustrates artificial approaches to sculpt DNA geometry with functional consequences. Similar approaches may be applicable to tune the stability of small DNA loops in eukaryotes.

biochemistry↗