Consistent with this conclusion, CD4+ T cells from these mice responded normally to activation gene in a temporally controlled manner  using a tamoxifen-regulated Cre recombinase. activation marker expression on CD4+ and CD8+ cells from GIMAP6fl/flCD2Cre and GIMAP6fl/fl mice. (TIF) pone.0196504.s008.TIF (127K) GUID:?13295035-04F9-432A-84F3-310AC3937B6F S9 Fig: T-cell dependent generation of antibody responses in GIMAP6fl/flCD2Cre mice. (TIF) pone.0196504.s009.TIF (91K) GUID:?466213E4-D287-4E0D-A006-786667AD9A48 Data Availability StatementAll relevant data are within the paper and its Supporting Information files. Abstract The GTPases of the immunity-associated proteins (GIMAP) GTPases are a family of proteins expressed strongly in the adaptive immune system. We have previously reported that in human cells one member of this family, GIMAP6, interacts with the ATG8 family member GABARAPL2, and is recruited to autophagosomes upon starvation, suggesting a role for GIMAP6 in the autophagic process. To study this possibility and the function of GIMAP6 in the immune system, we have established a mouse collection in which the gene can be inactivated by Cre-mediated recombination. In mice bred to carry the CD2Cre transgene such that the gene was deleted within the T and B cell lineages there was a 50C70% reduction in peripheral AR-42 (HDAC-42) CD4+ and CD8+ T cells. Analysis of splenocyte-derived proteins from these mice indicated increased levels of MAP1LC3B, particularly the lipidated LC3-II form, and S405-phosphorylation of SQSTM1. Electron microscopic measurements of CD4+ T cells indicated an increased mitochondrial/cytoplasmic volume ratio and increased numbers of autophagosomes. These results are consistent with autophagic disruption in the cells. However, T cells were largely normal in character, could be effectively activated and supported T cell-dependent antibody production. Treatment of CD4+ splenocytes from GIMAP6fl/flERT2Cre mice with 4-hydroxytamoxifen resulted in the disappearance of GIMAP6 within five days. In parallel, increased phosphorylation of SQSTM1 and TBK1 was observed. These results indicate a requirement for GIMAP6 in the maintenance of a normal peripheral adaptive immune system and a significant role for the protein in normal autophagic processes. Moreover, AR-42 (HDAC-42) as GIMAP6 is usually expressed in a cell-selective manner, this indicates the potential existence of a cell-restricted mode of autophagic regulation. Introduction The AIG1 family of GTPases are a group of proteins found sporadically in various eukaryotic phyla . The first member of the family, termed AIG1 (avrRpt2-induced gene 1), was recognized in the herb species contamination . In addition to plants, members of the family have also been recognized in a restricted quantity of other groups including protists , coral  and molluscs [5,6], (but not in e.g. or by contamination, in both coral  and molluscs [5,6] SAV1 AIG1 family members are also induced by pathogenic challenge, suggesting that they may have a significant role in conferring resistance to contamination. The link to host defence is usually further conserved in vertebrates, where the AIG1 family of GTPases is usually represented by the GIMAP family of proteins and expression of these is usually most prominent in cells of the adaptive immune system . In mammals, the GIMAP family AR-42 (HDAC-42) comprises 7C8 users (species-dependent) which are closely linked at a single locus (chromosome 7 in human, 6 in mouse) . The family can be split into two groups, depending on the presence or absence of membrane-anchoring domains. In mouse, GIMAPs 1, 3 and 5 are membrane-anchored, whereas GIMAPs 4, 6, 7, 8 and 9 are soluble proteins (observe ). In structural terms, GIMAPs have been placed in the non-Ras class of G proteins alongside septins and dynamins with which they share mechanisms of GTPase activation via molecular dimerization (including heterologous interactions within the GIMAP family). A role in molecular scaffold formation on intracellular membranes has been proposed . Historically, GIMAP5 has attracted the most research attention. Non-functional mutations of in both rats and mice are associated with severe peripheral T cell lymphopenia and increased susceptibility to autoimmune conditions such as type 1 diabetes and inflammatory bowel disease [11C16]. Interestingly, targeted mutations launched into the genes AR-42 (HDAC-42) encoding other GIMAP membrane-anchored proteins present in mice, namely GIMAP1  and GIMAP3 , also have an impact on T cell phenotypes, although that associated AR-42 (HDAC-42) with GIMAP3 is only seen clearly in the presence of an additional mutation . In contrast to what has been seen with the membrane-anchored proteins, mice transporting targeted mutations of the genes encoding two of the soluble GIMAPs show no (GIMAP4)  or very minor (GIMAP8)  changes in the lymphocyte populations, although T cells isolated from both of these mouse strains show altered kinetics of apoptosis [9,19]. We have previously reported work on another soluble member of the family, GIMAP6. We showed that human GIMAP6 interacts strongly with the autophagy-related ATG8 protein GABARAPL2 in transfected cell lines , an effect subsequently confirmed in a high-throughput screening assay . Moreover, GIMAP6 was recruited to autophagosomes upon activation of autophagy , perhaps via its conversation with GABARAPL2. This suggested either that GIMAP6 was selectively degraded by the autophagic pathway or, more interestingly, that it played a functional.