Elimination of GPI2 suppresses glycosylphosphatidylinositol GlcNAc transferase activity and alters GPI glycan modification in Trypanosoma brucei

Abstract

This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Identification of TbPBN1 in Trypanosoma brucei reveals a conserved heterodimeric architecture for glycosylphosphatidylinositol‐mannosyltransferase‐I

“…The TbGPI14 null parasites were viable but grew almost 2× more slowly than the parental cells (Figure 1a ). Reduced growth in culture has also been reported in T. brucei procyclic forms lacking other GPI synthesis enzymes (Hong et al., 2006 ; Jenni et al., 2021 ; Nagamune et al., 2000 , 2003 ; Verchère et al., 2021 ).…”

Persistence of Trypanosoma brucei as early procyclic forms and social motility are dependent on glycosylphosphatidylinositol transamidase

“…We recently reported that T. brucei procyclic forms lacking Tb GPI2 showed decreased viability on agarose plates (Jenni et al., 2021 ). To investigate whether Tb GPI8 was also required for survival on agarose plates, we determined the cell number and PDT in the presence and absence of tetracycline (Figure S5 , upper panels).…”

“…In all three GPI mutants, the surface is devoid of EP and GPEET procyclins (Güther et al., 2006 ; Lillico et al., 2003 ; Nagamune et al., 2004 ). As also seen with a procyclin null mutant (Vassella et al., 2003 ), deletion of Tb GPI8 (Lillico et al., 2003 ), Tb GPI10 (Nagamune et al., 2004 ) or Tb GPI2 (Jenni et al., 2021 ) in procyclic forms causes the accumulation of free (non‐protein‐linked) GPIs on the surface. This was not the case in Tb GPI12 null mutants, which did not express either procyclins or free GPIs on their surface (Güther et al., 2006 ).…”

“…Alexa Fluor 488‐conjugated or Cy3‐conjugated goat IgG secondary antibodies (Invitrogen, Thermo Fisher Scientific, 2 mg/ml) were diluted at 1:1,000. Flow cytometry of live trypanosomes was performed as described (Jenni et al., 2021 ). Parasites (4 × 10 6 cells) were harvested by centrifugation.…”

StaR-related lipid transfer-like domain-containing protein CLDP43 affects cardiolipin synthesis and mitochondrial function in Trypanosoma brucei

“…The cell suspension was spread on a microscopy slide and left to adhere for at least 10 min. Trypanosomes were fixed in 4% (w/v) paraformaldehyde in 1x PBS for 10 min [52]. The slides were washed 3 times with PBS for 5 min each and subsequently permeabilized in 0.2% (w/v) Triton X-100 in PBS for 5 min.…”