In this context, we have previously shown than the spacing of palmitates is key for fine-tuning protein localization. Moving the distal palmitoylation cysteine away from the isoprenylation site in a chimeric construct reduces its accummulation in MVB in response to alterations in lipid dynamics. In Aspergillus nidulans, both the putative Rho2-like protein and GFP-8 would be expected to be geranylgeranylated and palmitoylated, although, to the best of our knowledge, a homolog for GGTase I has not been biochemically identified in this organism and the information available on potential geranylgeranyltransferases only stems from sequence homology analysis on database searches. It can be noted that CINCCKVL chimeras displayed some degree of plasma Torin 1 membrane localization, although it was dependent on the cell type under study and on the construct used, with GFP-8 decorating the plasma membrane to a higher extent than tRFP-T-8. Nevertheless, the most consistent localization of CINCCKVL chimeric proteins, when lipidated, is the endolysosomal compartment. It could be hypothesized that the specific structure formed upon CINCCKVL lipidation promotes the interaction with specific microdomains that require the presence of cholesterol. Indeed, domains rich in free cholesterol have been described in MVB of endothelial cells under particular cell culture conditions. Moreover, the presence of lipid microdomains that segregate proteins in the yeast vacuole has been recently demonstrated in Saccharomyces cerevisiae. One of the most striking conclusions of this work is that the CINCCKVL sequence takes a route to endolysosomes present in the lower species but not used, to the best of our knowledge, by their endogenous proteins. Our findings may motivate the search for proteins with analogous sequences that may be sorted through this pathway, as well as the use of cells from lower species to completely unveil the sorting mechanism. This information will allow elucidating whether this sequence has evolved in the higher species to ensure endolysosomal delivery of specific proteins. In summary, our observations support the interpretation that the structures generated by precise lipidation sequences, and in particular by the CINCCKVL motif, may specifically interact with defined membrane components for protein targeting and sorting. These findings may set the basis for exploring the sorting of lipidated sequences to specific microdomains involved in MVB biogenesis in diverse species. Two-component systems are the major group of signal transduction systems that allow bacteria to cope with environmental changes. The classical two-component system is composed of a sensor kinase and a response regulator. Upon stimulation, the SK becomes autophosphorylated and transfers the phosphoryl group to its cognate RR, which modulates the response. Without stimulation, the response is terminated by dephosphorylation of the RR by either intrinsic activity or by the SK.