The guanine nucleotide exchange factor GBF1 participates in rotavirus replication

José L. Martínez; Francesca Arnoldi; Elisabeth M. Schraner; Catherine Eichwald; Daniela Silva-Ayala; Eunjoo Lee; Elizabeth Sztul; Óscar R. Burrone; Susana López; Carlos F. Arias

The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. show that this deletion renders GBF1 unable to support rotavirus replication in the presence 447 of BFA. It is possible that this 37 amino acid stretch could bind a rotaviral protein, although 448 it may also interact with a cellular factor to support virus replication. In fact, it has been 449 reported that Rab1b is able to interact with the N-terminus of GBF1 to modulate GBF1 450 function in the secretory pathway (85). Further experiments will have to be carried out to 451 explore more in detail the mechanism through which GBF1 supports rotavirus replication. The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. . https://doi.org/10.1101/619924 doi: bioRxiv preprint h at 37°C. The unbound virus was removed, and the cells were incubated for 16 h at 37 °C. 464 Finally, the cells were lysed by two cycles of freeze-thawing and the cell debris were removed 465 by centrifugation. The resulting viral lysate was stored at -70°C. The cell lysate was extracted with trichloromonofluoromethane as described above, and the 527 aqueous phase was mixed with 2.2 g of CsCl and TNC buffer up to 5 ml. The samples were 528 centrifuged at 158,000 x g for 18h at 4°C on a SW55Ti rotor (Beckman). Finally, the 529 opalescent bands corresponding to TLPs and DLPs were collected by puncture with a syringe 530 and stored at 4°C. Before use, the viral particles were desalted in a Sephadex G25 spin 531 column. 532 All rights reserved. No reuse allowed without permission.