{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T12:01:01Z","timestamp":1787313661842,"version":"build-2736575974"},"reference-count":41,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T00:00:00Z","timestamp":1774483200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Institute of Biochemistry and Biophysics Polish Academy of Sciences"},{"id":[{"id":"https:\/\/ror.org\/034tvp782","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoF"],"abstract":"<jats:p>The Rot1 protein is a chaperone involved in glycosylation, dolichol phosphorylation, cell wall synthesis, and protein folding in the yeast Saccharomyces cerevisiae. Available information on cell wall defects in the S. cerevisiae rot1-1 mutant and the association of Rot1 with protein glycosylation suggest that in the case of Candida albicans, Rot1 may be involved in pathogenesis, since both cell wall synthesis and protein glycosylation are closely related to the formation of pathogenic structures in C. albicans. As Rot1 has not been found in humans, it seems particularly attractive for study in the context of C. albicans pathogenicity. This protein takes on additional significance because deletion of the gene that encodes Rot1 is lethal for yeast. In this study, we cloned and analyzed the function of the candidate protein CaRot1 from C. albicans in the S. cerevisiae rot1\u0394\/ROT1 mutant. Furthermore, we investigated the consequences of restricted CaROT1 expression in C. albicans. We have shown that a low amount of Rot1 limits the transfer of oligosaccharide to protein, inhibits the activity of the first steps of oligosaccharide formation on dolichyl diphosphate, changes the composition of the cell wall, limits the protection of C. albicans against ER and abiotic stress, and finally prevents filamentation, which is an invasive structure of C. albicans.<\/jats:p>","DOI":"10.3390\/jof12040244","type":"journal-article","created":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T13:50:15Z","timestamp":1774533015000},"page":"244","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["The Essential Rot1 Protein Links Glycosylation, Cell Wall Integrity, and Pathogenic Development in Candida albicans"],"prefix":"10.3390","volume":"12","author":[{"given":"Anna","family":"Janik","sequence":"first","affiliation":[{"name":"Institute of Biochemistry and Biophysics Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ewa","family":"Zatorska","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry and Biophysics Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Urszula","family":"Perli\u0144ska-Lenart","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry and Biophysics Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9279-0577","authenticated-orcid":false,"given":"Sebastian","family":"Pi\u0142syk","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry and Biophysics Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2012-7781","authenticated-orcid":false,"given":"Joanna S.","family":"Kruszewska","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry and Biophysics Polish Academy of Sciences, Pawinskiego 5a, 02-106 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"3271","DOI":"10.1101\/gad.13.24.3271","article-title":"The TOR signaling cascade regulates gene expression in response to nutrients","volume":"13","author":"Cardenas","year":"1999","journal-title":"Genes Dev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2235","DOI":"10.1093\/emboj\/17.8.2235","article-title":"Cell wall integrity modulates RHO1 activity via the exchange factor ROM2","volume":"17","author":"Bickle","year":"1998","journal-title":"EMBO J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"597","DOI":"10.1093\/jb\/mvj063","article-title":"Saccharomyces cerevisiae Rot1p is an ER-localized membrane protein that may function with BiP\/Kar2p in protein folding","volume":"139","author":"Takeuchi","year":"2006","journal-title":"J. Biochem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2390","DOI":"10.1242\/jcs.002758","article-title":"Rot1 plays an antagonistic role to Clb2 in actin cytoskeleton dynamics throughout the cell cycle","volume":"120","author":"Juanes","year":"2007","journal-title":"J. Cell Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3514","DOI":"10.1091\/mbc.e07-12-1289","article-title":"Saccharomyces cerevisiae Rot1 is an essential molecular chaperone in the endoplasmic reticulum","volume":"19","author":"Takeuchi","year":"2008","journal-title":"Mol. Biol. Cell"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1002\/yea.1479","article-title":"Dissecting the role of dolichol in cell wall assembly in the yeast mutants impaired in early glycosylation reactions","volume":"24","author":"Janik","year":"2007","journal-title":"Yeast"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1093\/glycob\/cws068","article-title":"The essential endoplasmic reticulum chaperone Rot1 is required for protein N- and O-glycosylation in yeast","volume":"22","author":"Pasikowska","year":"2012","journal-title":"Glycobiology"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"6564","DOI":"10.1016\/j.febslet.2005.10.063","article-title":"Yeast oligosaccharyltransferase consists of two functionally distinct sub-complexes, specified by either the Ost3p or Ost6p subunit","volume":"579","author":"Schwarz","year":"2005","journal-title":"FEBS Lett."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1396","DOI":"10.1093\/glycob\/cwj025","article-title":"The 3. 4-kDa Ost4 protein is required for the assembly of two distinct oligosaccharyltransferase complexes in yeast","volume":"15","author":"Spirig","year":"2005","journal-title":"Glycobiology"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1604","DOI":"10.1093\/glycob\/cwab084","article-title":"Functional analysis of Ost3p and Ost6p containing yeast oligosaccharyltransferases","volume":"31","author":"Neuhaus","year":"2021","journal-title":"Glycobiology"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"100809","DOI":"10.1016\/j.jbc.2021.100809","article-title":"Substrate specificities and reaction kinetics of the yeast oligosaccharyltransferase isoforms","volume":"296","author":"Eyring","year":"2021","journal-title":"J. Biol. Chem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"628","DOI":"10.1038\/sj.embor.embor864","article-title":"O-mannosylation precedes and potentially controls the N-glycosylation of a yeast cell wall glycoprotein","volume":"4","author":"Ecker","year":"2003","journal-title":"EMBO Rep."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"3163","DOI":"10.1099\/mic.0.27292-0","article-title":"Rot1p of Saccharomyces cerevisiae is a putative membrane protein required for normal levels of the cell wall 1,6-\u03b2-glucan","volume":"150","author":"Machi","year":"2004","journal-title":"Microbiology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.fgb.2014.05.004","article-title":"Candida albicans cis-prenyltransferase Rer2 is required for protein glycosylation, cell wall integrity and hyphae formation","volume":"69","author":"Juchimiuk","year":"2014","journal-title":"Fungal Genet. Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2265","DOI":"10.1016\/j.bbagen.2015.08.012","article-title":"Dolichol phosphate mannose synthase from the pathogenic yeast Candida albicans is a multimeric enzyme","volume":"1850","author":"Juchimiuk","year":"2015","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"789","DOI":"10.1016\/j.bbagen.2017.01.019","article-title":"The role of Alg13 N-acetylglucosaminyl transferase in the expression of pathogenic features of Candida albicans","volume":"1861","author":"Niewiadomska","year":"2017","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Janik, A., Niewiadomska, M., Perlinska-Lenart, U., Lenart, J., Ko\u0142akowski, D., Skorupinska-Tudek, K., Swiezewska, E., Kruszewska, J.S., and Palamarczyk, G. (2019). Inhibition of dephosphorylation of dolichyl diphosphate alters the synthesis of dolichol and hinders protein N-glycosylation and morphological transitions in Candida albicans. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20205067"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"5809","DOI":"10.1016\/j.jmb.2020.09.003","article-title":"A single site phosphorylation on Hsp82 ensures cell survival during starvation in Saccharomyces cerevisiae","volume":"432","author":"Shang","year":"2020","journal-title":"J. Mol. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1007\/BF00318659","article-title":"One-step transformation of yeast in stationary phase","volume":"21","author":"Chen","year":"1992","journal-title":"Curr. Genet."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1093\/genetics\/134.3.717","article-title":"Isogenic strain construction and gene mapping in Candida albicans","volume":"134","author":"Fonzi","year":"1993","journal-title":"Genetics"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1016\/0003-2697(87)90021-2","article-title":"Single-step method of RNA isolation by acid guanidinum thiocyanate-phenol-chloroform extraction","volume":"162","author":"Chomczynski","year":"1987","journal-title":"Anal. Biochem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"E36","DOI":"10.1093\/nar\/30.9.e36","article-title":"Relative Expression Software Tool (REST\u00a9) for group wise comparison and statistical analysis of relative expression results in real-time PCR","volume":"30","author":"Pfaffl","year":"2002","journal-title":"Nucleic Acids Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1042\/bj1480245","article-title":"The formation of mono-N-acetylhexosamine derivatives of dolichol diphosphate by pig liver microsomal fractions","volume":"148","author":"Palmarczyk","year":"1975","journal-title":"Biochem. J."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1074\/jbc.M510360200","article-title":"Outer chain-glycans are required for cell wall integrity and virulence of Candida albicans","volume":"281","author":"Bates","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1556\/ABiol.52.2001.2-3.12","article-title":"Cell wall composition and polysaccharide synthase activity changes following photoinduction in Trichoderma viride","volume":"52","author":"Nemcovic","year":"2001","journal-title":"Acta Biol. Hung."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1006\/abio.1999.4294","article-title":"A single method for carbohydrate and protein contents in glycoprotein bands separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis","volume":"275","author":"Zdebska","year":"1999","journal-title":"Anal. Biochem."},{"key":"ref_27","first-page":"350","article-title":"Colorimetric method for determination of sugar and related substrates","volume":"28","author":"Dubois","year":"1956","journal-title":"Anal. Biochem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"886","DOI":"10.1128\/EC.2.5.886-900.2003","article-title":"Chitin synthesis in Saccharomyces cerevisiae in response to supplementation of growth medium with glucosamine and cell stress","volume":"2","author":"Bulik","year":"2003","journal-title":"Eucaryotic Cell"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1543","DOI":"10.1099\/13500872-140-7-1543","article-title":"Purification and characterization of two forms of N-acetylglucosaminidase from Candida albicans showing widely different outer chain glycosylation","volume":"140","author":"Molloy","year":"1994","journal-title":"Microbiology"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2186","DOI":"10.1021\/bi00578a008","article-title":"Mechanism of action of tunicamycin on the UDP-GlcNAc: Dolichyl-phosphate GlcNAc-1-phosphate transferase","volume":"18","author":"Heifetz","year":"1979","journal-title":"Biochemistry"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"41455","DOI":"10.1074\/jbc.M105544200","article-title":"The CWH8 gene encodes a dolichyl pyrophosphate phosphatase with luminal orientated active site in the endoplasmic reticulum of Saccharomyces cerevisiae","volume":"276","author":"Fernandez","year":"2001","journal-title":"J. Biol. Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"6295","DOI":"10.1038\/s41467-025-61678-1","article-title":"Distinct echinocandin responses of Candida albicans and Candida auris cell walls revealed by solid-state NMR","volume":"16","author":"Widanage","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1038\/nrmicro2711","article-title":"Candida albicans morphogenesis and host defence: Discriminating invasion from colonization","volume":"10","author":"Gow","year":"2013","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"406","DOI":"10.1016\/j.mib.2012.04.005","article-title":"Importance of the Candida albicans cell wall during commensalism and infection","volume":"15","author":"Gow","year":"2012","journal-title":"Curr. Opin. Microbiol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Chow, E.W.L., Pangh, L.M., and Wang, Y. (2021). From Jekyll to Hyde: The yeast\u2013hyphal transition of Candida albicans. Pathogens, 10.","DOI":"10.20944\/preprints202106.0386.v1"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1128","DOI":"10.1128\/microbiolspec.FUNK-0035-2016","article-title":"The fungal cell wall: Structure, biosynthesis, and function","volume":"5","author":"Gow","year":"2017","journal-title":"Microbiol. Spectr."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"RP100569","DOI":"10.7554\/eLife.100569.3","article-title":"\u03b2-1,6- Glucan plays a central role in the structure and remodeling of the bilaminate fungal cell wall","volume":"13","author":"Bekirian","year":"2024","journal-title":"eLife"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1016\/j.ijantimicag.2009.04.011","article-title":"In vitro analyses of the combination of high-dose doxycycline and antifungal agents against Candida albicans biofilms","volume":"34","author":"Miceli","year":"2009","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1093\/jac\/dkg117","article-title":"Caspofungin: The first representative of a new antifungal class","volume":"51","author":"Herbrecht","year":"2003","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1071","DOI":"10.1128\/AAC.00946-13","article-title":"Paradoxical growth of Candida albicans in the presence of caspofungin is associated with multiple cell wall rearrangements and decreased virulence","volume":"58","author":"Rueda","year":"2014","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Toyoda, Y., and Saitoh, S. (2021). Fission yeast TORC2 signaling pathway ensures cell proliferation under glucose-limited, nitrogen-replete conditions. Biomolecules, 11.","DOI":"10.3390\/biom11101465"}],"container-title":["Journal of Fungi"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2309-608X\/12\/4\/244\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,1]],"date-time":"2026-04-01T16:13:22Z","timestamp":1775060002000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2309-608X\/12\/4\/244"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,26]]},"references-count":41,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["jof12040244"],"URL":"https:\/\/doi.org\/10.3390\/jof12040244","relation":{},"ISSN":["2309-608X"],"issn-type":[{"value":"2309-608X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,26]]}}}