title: Identification of Haloferax volcanii Pilin N-Glycans with Diverse Roles in Pilus Biosynthesis, Adhesion, and Microcolony Formation.
abstract: N-Glycosylation is a post-translational modification common to all three domains of life. In many archaea, the oligosacharyltransferase (AglB)-dependent N-glycosylation of flagellins is required for flagella assembly. However, whether N-glycosylation is required for the assembly and/or function of the structurally related archaeal type IV pili is unknown. Here, we show that of six Haloferax volcanii adhesion pilins, PilA1 and PilA2, the most abundant pilins in pili of wild-type and ΔaglB strains, are modified under planktonic conditions in an AglB-dependent manner by the same pentasaccharide detected on H. volcanii flagellins. However, unlike wild-type cells, which have surfaces decorated with discrete pili and form a dispersed layer of cells on a plastic surface, ΔaglB cells have thick pili bundles and form microcolonies. Moreover, expressing PilA1, PilA2, or PilA6 in ΔpilA[1-6]ΔaglB stimulates microcolony formation compared with their expression in ΔpilA[1-6]. Conversely, expressing PilA3 or PilA4 in ΔpilA[1-6] cells results in strong surface adhesion, but not microcolony formation, and neither pilin stimulates surface adhesion in ΔpilA[1-6]ΔaglB cells. Although PilA4 assembles into pili in the ΔpilA[1-6]ΔaglB cells, these pili are, unlike wild-type pili, curled, perhaps rendering them non-functional. To our knowledge, this is the first demonstration of a differential effect of glycosylation on pilus assembly and function of paralogous pilins. The growth of wild-type cells in low salt media, a condition that decreases AglB glycosylation, also stimulates microcolony formation and inhibits motility, supporting our hypothesis that N-glycosylation plays an important role in regulating the transition between planktonic to sessile cell states as a response to stress.
authors.affiliation: From the Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19104 and.
authors.forename: Rianne N
authors.initials: RN
authors.lastname: Esquivel
authors.affiliation: the Institute of Plant Biology and Biotechnology, University of Münster, Münster 48143, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Schulze
authors.affiliation: From the Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19104 and.
authors.forename: Rachel
authors.initials: R
authors.lastname: Xu
authors.affiliation: the Institute of Plant Biology and Biotechnology, University of Münster, Münster 48143, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Hippler
authors.affiliation: From the Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19104 and pohlschr@sas.upenn.edu.
authors.forename: Mechthild
authors.initials: M
authors.lastname: Pohlschroder
mesh_terms: D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002448:Cell Adhesion / Q000502:physiology; D036781:Fimbriae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010861:Fimbriae, Bacterial / Q000378:metabolism / Q000648:ultrastructure; D006031:Glycosylation; D019622:Haloferax volcanii / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism; D055503:Protein Multimerization; D011499:Protein Processing, Post-Translational; D053719:Tandem Mass Spectrometry
chemical_list: D019843:Archaeal Proteins; D011134:Polysaccharides; D036781:Fimbriae Proteins
keywords: Haloferax volcanii; N-linked glycosylation; adhesion; archaea; biofilm; flagella; flagellin; pili; pilin; type IV pili