RT Journal Article
SR Electronic
T1 Intrabody induced cell death by targeting the T. brucei cytoskeletal protein TbBILBO1
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2021.07.18.452872
DO 10.1101/2021.07.18.452872
A1 Christine E. Broster Reix
A1 Miharisoa Rijatiana Ramanantsalama
A1 Carmelo Di Primo
A1 Laëtitia Minder
A1 Mélanie Bonhivers
A1 Denis Dacheux
A1 Derrick R. Robinson
YR 2021
UL http://biorxiv.org/content/early/2021/07/20/2021.07.18.452872.abstract
AB Trypanosoma brucei belongs to a genus of protists that cause life-threatening and economically important diseases of human and animal populations in Sub-Saharan Africa. T. brucei cells are covered in surface glycoproteins some of which are used to escape the host immune system. Exo-/endocytotic trafficking of these and other molecules occurs via a single copy organelle called the flagellar pocket (FP). The FP is maintained and enclosed around the flagellum by the flagellar pocket collar (FPC). To date, the most important cytoskeletal component of the FPC is an essential, calcium-binding, polymer-forming protein called TbBILBO1. In searching for novel immune-tools to study this protein, we raised nanobodies against TbBILBO1. Nanobodies (Nb) that were selected according to their binding properties to TbBILBO1, were tested as immunofluorescence tools, and expressed as intrabodies (INb). One of them, Nb48, proved to be the most robust nanobody and intrabody. We further demonstrate that inducible, cytoplasmic expression of INb48 was lethal to these parasites, producing abnormal phenotypes resembling those of TbBILBO1 RNAi knockdown. Our results validate the feasibility of generating functional single-domain antibody derived intrabodies to target trypanosome cytoskeleton proteins.