Abstract

Biofilms are the predominant lifestyle of microorganisms and offer protection against environmental stress. Archaeal biofilm development remains poorly understood relative to its bacterial counterpart, and the fundamental differences between these two domains prevent direct extrapolation from bacterial models. To address this gap, we used quantitative proteomics and transcriptomics to characterize the molecular changes accompanying biofilm formation and maturation in the model archaeon Haloferax volcanii. Protein and transcript abundance were measured across seven conditions spanning both lifestyles and multiple timepoints, with glycosylated proteoforms quantified as entities distinct from their unmodified counterparts, covering both established biofilm-associated structures and newly identified candidates. Known biofilm-associated type IV pili showed coordinated behavior across the time course, while archaella demonstrated retention and accumulation in biofilm samples. The two N-glycosylation pathways diverged from one another, with Agl15-dependent proteins enriched in planktonic cells while AglB-dependent proteins changed little, even as their AglB-dependent glycan products were among the most strongly biofilm enriched features detected. Clustering analysis also identified numerous annotated proteins, hypothetical proteins, and probable glycoproteins whose abundances correlated with groups we termed biofilm adherence and biofilm maturation. Together, these findings provide a quantitative, glycosylation-informed molecular basis for archaeal biofilm development, and define priority targets for future functional studies.

Publication Date

8-3-2026

Document Type

Thesis

Student Type

Graduate

Degree Name

Bioinformatics (MS)

College

College of Science

Advisor

Stefan Schulze

Advisor/Committee Member

Crista Wadsworth

Advisor/Committee Member

Robert Osgood

Campus

RIT – Main Campus

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