O-fucosylated glycoproteins form assemblies in close proximity to the nuclear pore complexes of Toxoplasma gondii.
Toxoplasma gondii is an intracellular parasite that causes disseminated infections in fetuses and immunocompromised individuals. Although gene regulation is important for parasite differentiation and pathogenesis, little is known about protein organization in the nucleus. Here we show that the fucose-binding Aleuria aurantia lectin (AAL) binds to numerous punctate structures in the nuclei of tachyzoites, bradyzoites, and sporozoites but not oocysts. AAL also binds to Hammondia and Neospora nuclei but not to more distantly related apicomplexans. Analyses of the AAL-enriched fraction indicate that AAL binds O-linked fucose added to Ser/Thr residues present in or adjacent to Ser-rich domains (SRDs). Sixty-nine Ser-rich proteins were reproducibly enriched with AAL, including nucleoporins, mRNA-processing enzymes, and cell-signaling proteins. Two endogenous SRDs-containing proteins and an SRD-YFP fusion localize with AAL to the nuclear membrane. Superresolution microscopy showed that the majority of the AAL signal localizes in proximity to nuclear pore complexes. Host cells modify secreted proteins with O-fucose; here we describe the O-fucosylation pathway in the nucleocytosol of a eukaryote. Furthermore, these results suggest O-fucosylation is a mechanism by which proteins involved in gene expression accumulate near the NPC.
Department of Molecular and Cell Biology, Boston University Goldman School of Dental Medicine, Boston, MA 02118.
Giulia
G
Bandini
Department of Molecular and Cell Biology, Boston University Goldman School of Dental Medicine, Boston, MA 02118; Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118.
John R
JR
Haserick
Department of Molecular and Cell Biology, Boston University Goldman School of Dental Medicine, Boston, MA 02118; Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118.
Edwin
E
Motari
Department of Biology, University of Pennsylvania, Philadelphia, PA 19104.
Dinkorma T
DT
Ouologuem
Whitehead Institute for Biomedical Research, Cambridge, MA 02142.
Sebastian
0000-0002-5237-1095
S
Lourido
Department of Biology, University of Pennsylvania, Philadelphia, PA 19104.
David S
DS
Roos
Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118.
Catherine E
CE
Costello
Department of Molecular and Cell Biology, Boston University Goldman School of Dental Medicine, Boston, MA 02118; robbinspw2@gmail.com jsamuels@bu.edu.
Phillips W
PW
Robbins
Department of Molecular and Cell Biology, Boston University Goldman School of Dental Medicine, Boston, MA 02118; robbinspw2@gmail.com jsamuels@bu.edu.
John
J
Samuelson
D000595:Amino Acid Sequence; D000818:Animals; D002453:Cell Cycle; D002460:Cell Line; D005643:Fucose / Q000378:metabolism*; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D037102:Lectins / Q000378:metabolism; D051379:Mice; D009685:Nuclear Envelope / Q000378:metabolism; D022022:Nuclear Pore / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism; D000072417:Protein Domains; D015800:Protozoan Proteins / Q000737:chemistry / Q000378:metabolism*; D013045:Species Specificity; D014122:Toxoplasma / Q000378:metabolism*
D006023:Glycoproteins; D037102:Lectins; D011134:Polysaccharides; D015800:Protozoan Proteins; C064549:lectin, Aleuria aurantia; D005643:Fucose
fucose; nuclear glycosylation; nuclear pore complex; toxoplasma
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