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Gill, V.

Publications and source records attributed to Gill, V..

3 recordsLinked to original sources

Characterizing the Prey Landscape of Endangered Cook Inlet Beluga Whales Using an eDNA Time Series

Our ability to understand habitat utilization in the context of foraging, prey dynamics, and anthropogenic disturbance is critical for the effective management of protected species globally. Here we examine the occurrence of the endangered Cook Inlet beluga whale (Delphinapterus leucas) in the context of seasonally changing prey fields by leveraging quantitative eDNA metabarcoding approaches. Specifically, eDNA approaches were used to generate estimates of fish abundances using a joint modeling framework that derives total DNA concentration from a coho (Oncorhynchus kisutch) qPCR assay and bias-corrected proportional abundance of multiple salmonid species using MiFish 12S metabarcoding of environmental and mock community samples. Temporal patterns of beluga whale occurrence were documented in two estuarine systems using visual observations, metabarcoding, and a novel beluga whale qPCR assay. We demonstrate that weekly time series before, during, and after beluga whale occurrence provide sufficient resolution to resolve changes in predator occurrence and prey field shifts across Cook Inlet. Specifically, we observed Cook Inlet beluga whales co-occurring with increases in coho DNA concentrations in the vicinity of Twentymile River at the terminus of Turnagain Arm. Monthly sampling on the Kenai River captured known seasonal anadromous fish migration patterns and identified a clear mismatch between Cook Inlet beluga whale habitat utilization and spring eulachon (Thaleichthys pacificus) runs. Our results also provide valuable insights to improve quantitative metabarcoding approaches through informed selection of qPCR targets and mock communities to support future eDNA-based studies in the highly turbid, dynamic habitats of Cook Inlet. Specifically, our results highlight the need for increased temporal sampling frequency as opposed to additional spatial and depth sampling in systems with extreme tides, as well as the use of larger pore sizes (> 5um) to facilitate filtration in samples with high suspended glacial silt loads. We also document the capacity for eDNA signatures to integrate across 18 km spatial scales during exceptionally large tidal bores (> 10 m) that characterize Turnagain Arm. Ultimately, this study demonstrates the value of pairing visual surveys of endangered Cook Inlet beluga whales with strategic shore-based eDNA sampling to generate information on the co-occurrence of Cook Inlet beluga whales and putative prey fields.

ecology↗

Generative Design of New-to-nature Biosynthetic Assembly Lines with Genomic Language Modeling

Reprogramming biosynthetic assembly lines can extend biosynthesis beyond the chemical space explored by nature. However, this remains difficult because assembly-line function depends on coordinated interactions across large multidomain enzymes. Here, we couple gLM2, a genomic language model trained on metagenomic sequences, with discrete diffusion and domain-level conditioning to enable generative design and optimization of biosynthetic gene clusters. We apply this approach to a chimeric type I polyketide synthase (PKS) engineered to produce {delta}-valerolactam, a molecule not naturally synthesized by PKSs. Through iterative redesign of two multi-domain regions in the context of the full PKS sequence, gLM2 progressively improved {delta}-valerolactam production, yielding variants with up to 9.4-fold higher titer than the starting enzyme. Together, these results demonstrate that evolutionary sequence information can be learned and applied to complex, multi-domain enzyme design problems, expanding biosynthetic assembly lines to produce molecules outside their natural biosynthetic repertoire.

bioengineering↗

The Structural Basis of alpha/beta-tubulin Assembly and Disassembly by Tubulin Cofactors

Microtubules polymerize from cytoplasmic pools of soluble {beta}-tubulin heterodimers that support diverse cellular functions. The tubulin cofactors, TBCC, TBCD, TBCE, and the Arl2 GTPase, form TBC-DEG assemblies that regulate {beta}-tubulin assembly and disassembly from - and {beta}-tubulins, yet their underlying mechanisms remain incompletely understood. Here, we reconstitute the human TBC-DE and TBC-DEG assemblies from eukaryotic cells co-purified with monomeric {beta}-tubulin intermediates and determine their cryo-EM structures. The structures reveal that TBC-DEG disassembles {beta}-tubulin by releasing -tubulin through a lever-arm-like rotation in TBCE coupled to major conformational change in Arl2 upon its nucleotide release, while TBCD tightly holds {beta}-tubulin. TBCD dissociates -tubulin by refolding the {beta}-tubulin H10-S8 loop at its intradimer interface. The TBC-DEG-{beta}-tubulin or TBC-DE-{beta}-tubulin assemblies undergo extensive back-to-back dimerization mediated by {beta}-{beta}-tubulin homodimers, formed through their dissociated H8 helices at unoccupied intradimer interfaces. Structural comparisons demonstrate that the TBCE mechanical rotation, driven by the Arl2 GTPase cycle, either delivers -tubulin or removes it from beneath the TBCD-bound {beta}-tubulin and is directionally regulated by TBCC stabilizing {beta}-tubulin interfaces. Our findings suggest that TBC-DEG/TBCC catalyzing heterodimerization of -tubulin with {beta}-tubulin may have evolved to counteract the {beta}-tubulin intrinsic tendency to form off-pathway toxic homodimers through its exposed -tubulin-binding intradimer interface.

biochemistry↗