The murine cytomegalovirus immunoevasin gp40/m152 inhibits NKG2D receptor RAE-1γ by intracellular retention and cell surface masking.

NKG2D (also known as KLRK1) is a crucial natural killer (NK) cell-activating receptor, and the murine cytomegalovirus (MCMV) employs multiple immunoevasins to avoid NKG2D-mediated activation. One of the MCMV immunoevasins, gp40 (m152), downregulates the cell surface NKG2D ligand RAE-1γ (also known as Raet1c) thus limiting NK cell activation. This study establishes the molecular mechanism by which gp40 retains RAE-1γ in the secretory pathway. Using flow cytometry and pulse-chase analysis, we demonstrate that gp40 retains RAE-1γ in the early secretory pathway, and that this effect depends on the binding of gp40 to a host protein, TMED10, a member of the p24 protein family. We also show that the TMED10-based retention mechanism can be saturated, and that gp40 has a backup mechanism as it masks RAE-1γ on the cell surface, blocking the interaction with the NKG2D receptor and thus NK cell activation.

Calibrating insect age at eclosion by size in a gregarious carrion beetle Thanatophilus sinuatus (Staphylinidae: Silphinae).

Recent discoveries have shown that the physiological age at eclosion of forensically useful beetles differs between males and females and between beetles of various sizes. Accordingly, it was postulated that the size and sex of the beetles at eclosion may be used to calibrate their age, which may improve the accuracy of age (and post-mortem interval) estimates in forensic entomology. In this study, we derived thermal summation models for the eclosion for the Central European population of carrion beetles Thanatophilus sinuatus (Fabricius, 1775), (Staphylinidae: Silphinae), and tested the usefulness of sex and size for the calibration of beetle age at eclosion. Although in previous developmental studies, the beetles were reared individually, we reared them in larval aggregations, since in natural conditions T. sinuatus beetles are gregarious. Weak (r2 between 5% and 13%) negative correlations were observed between the size and age of T. sinuatus males or females at eclosion, demonstrating that calibration of age by beetle size and sex may bring only minimal benefits regarding the accuracy of age estimation in this species. However, it may still be worthwhile in the case of extremely large or small beetles. Moreover, the total development times recorded in this study were much shorter than in the previous T. sinuatus study, at 14°C by about 15 days and at 26°C by about 2 days. These differences emphasise the importance of gregariousness for the development of carrion beetles, and at the same time highlight the need for the ecologically-relevant protocols of development studies in forensic entomology.

Competition, cooperation, and parental effects in larval aggregations formed on carrion by communally breeding beetles Necrodes littoralis (Staphylinidae: Silphinae).

Aggregations of juveniles are dominant forms of social life in some insect groups. Larval societies are shaped by competitive and cooperative interactions of the larvae, in parallel with parental effects. Colonies of necrophagous larvae are excellent systems to study these relationships. Necrodes littoralis (Staphylinidae: Silphinae), a carrion beetle that colonizes cadavers of large vertebrates, forms massive juvenile aggregations. By spreading over carrion anal and oral exudates, the beetles form the feeding matrix, in which the heat is produced and by which adults presumably affect the fitness of the larvae. We predict that exploitative competition shapes the behavior of N. littoralis larvae in their aggregations. However, cooperative interactions may also operate in these systems due mainly to the benefits of collective exodigestion. Moreover, indirect parental effects (i.e., formation of the feeding matrix) probably modulate larval interactions within the aggregations. By manipulating parental effects (present/absent) and larval density (0.02-1.9 larvae/g of meat), we found a strong negative group-size effect on fitness components of N. littoralis, in colonies with parental effects over almost the entire density range, and in colonies without parental effects for densities larger than 0.5 larva/g. This was accompanied by positive group-size effects in terms of development time (it shortened with larval density) and thermogenesis (it increased with larval density). A pronounced positive group-size effect on juvenile fitness was found only in colonies without parental effects and only in the low-density range. These results support the hypothesis that larval societies of N. littoralis are shaped by exploitation competition.

Cytomegalovirus gp40/m152 Uses TMED10 as ER Anchor to Retain MHC Class I.

The murine cytomegalovirus immunoevasin m152/gp40 binds major histocompatibility complex (MHC) class I molecules and retains them in the early secretory pathway by a previously unknown mechanism, preventing antigen presentation to CD8+ T cells. We show that retention of class I and of gp40 itself depends on a lumenal linker sequence in gp40. With unbiased co-immunoprecipitation and mass spectrometry, we find that, through this linker, gp40 binds to TMED10/Tmp21/p24δ1, a member of the p24 family of endoplasmic reticulum (ER)/Golgi transmembrane proteins. We show that the C-terminal KKxxx Golgi-to-ER retrieval signal of TMED10 is required for gp40-mediated retention of class I. We thus identify a viral interaction partner of the p24 proteins and their exploitation for viral immune evasion.