2020 Theses Doctoral

MID1IP1 and CCT2 in HIV-1 Transduction

Ermakova, Marina

HIV-1 completes its life cycle by coopting host proteins. Hundreds of proteins have been identified as potential host factors functioning in viral infection through screens, two of which are MID1IP1 and CCT2. Little is known about MID1IP1, but its localization to microtubules may suggest a cytoskeletal function and a possible role in microtubule transport of HIV-1 viral cores. We use the CRISPR/Cas9 system to create frameshift mutations in MID1IP1 in 293 cells and find that these mutations do not produce effects on HIV-1 transduction in experiments capable of assaying for completion of the life cycle from initial entry into host cells to gene expression. Furthermore, we were unable to find an effect on the staining for markers of microtubule stability using Western blots as a result of the mutations in these cells. CCT2 is a component of the TRiC/CCT protein folding complex whose substrates include actin and tubulin, which also suggests that CCT2 might function in the HIV-1 life cycle in a cytoskeleton-dependent manner. siRNA knockdowns in TE671 cells demonstrate a slight effect on HIV-1 transduction. Our data does not support a role for MID1IP1 in the entry stage of the HIV-1 life cycle, but does suggest CCT2 may be a potential candidate for further study.