HIV-1 Envelope Glycoprotein Trafficking through the Endosomal Recycling Compartment Is Required for Particle Incorporation

ABSTRACTThe human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein (Env) encodes specific trafficking signals within its long cytoplasmic tail (CT) that regulate incorporation into HIV-1 particles. Rab11-family interacting protein 1C (FIP1C) and Rab14 are host trafficking factors required for Env particle incorporation, suggesting that Env undergoes sorting from the endosomal recycling compartment (ERC) to the site of particle assembly on the plasma membrane. We disrupted outward sorting from the ERC by expressing a C-terminal fragment of FIP1C (FIP1C560–649) and examined the consequences on Env trafficking and incorporation into particles. FIP1C560–649reduced cell surface levels of Env and prevented its incorporation into HIV-1 particles. Remarkably, Env was trapped in an exaggerated perinuclear ERC in a CT-dependent manner. Mutation of either the Yxxϕ endocytic motif or the YW795motif in the CT prevented Env trapping in the ERC and restored incorporation into particles. In contrast, simian immunodeficiency virus SIVmac239 Env was not retained in the ERC, while substitution of the HIV-1 CT for the SIV CT resulted in SIV Env retention in this compartment. These results provide the first direct evidence that Env traffics through the ERC and support a model whereby HIV-1 Env is specifically targeted to the ERC prior to FIP1C- and CT-dependent outward sorting to the particle assembly site on the plasma membrane.IMPORTANCEThe HIV envelope protein is an essential component of the viral particle. While many aspects of envelope protein structure and function have been established, the pathway it follows in the cell prior to reaching the site of particle assembly is not well understood. The envelope protein has a very long cytoplasmic tail that interacts with the host cell trafficking machinery. Here, we utilized a truncated form of the trafficking adaptor FIP1C protein to arrest the intracellular transport of the envelope protein, demonstrating that it becomes trapped inside the cell within the endosomal recycling compartment. Intracellular trapping resulted in a loss of envelope protein on released particles and a corresponding loss of infectivity. Mutations of specific trafficking motifs in the envelope protein tail prevented its trapping in the recycling compartment. These results establish that trafficking to the endosomal recycling compartment is an essential step in HIV envelope protein particle incorporation.

Influence of Ti and Zn on Particle Incorporation of AlB2, B and B4C Particles in Aluminum Using the Stir Casting Process

The outstanding performance of many aluminum matrix composites (AMCs) regarding specific stiffness makes AMCs attractive materials for lightweight construction. Low density boride compounds promise both an increase in stiffness and decrease in composite density. Therefore for this study AlB2, B and B4C were chosen for composite manufacturing. The composites were fabricated with the stir casting process. To avoid gas entrapment during mixing and ensure nonporous composites, partial vacuum was adapted during particle feeding and stirring. Poor wettability of used particle material in contact with liquid aluminum hindered particle incorporation, but alloying elements such as titanium were shown to affect wettability and particle incorporation for B4C. Zn had no influence on wettability or reactivity and did not improve particle incorporation. In contrast to Zn, Ti improved adhesion and wettability, but particle incorporation was improved exclusively for B4C. Besides alloying Ti, the use of high-shear force mixers improved particle incorporation enabling uniform particle distribution. AMCs with up to 12 vol.% of B4C particles were produced via stir casting without alloying Ti.