HLA-DR histocompatibility antigens are assembled in the endoplasmic reticulum. This assembly has been studied in vitro and in vivo. Three polypeptides are involved in forming the oligomeric structure of HLA-DR antigens, DR alpha chains (molecular weight 35,000), DR beta chains (molecular weight 29,000) and DR gamma chains (molecular weight 33,000). They are cotranslationally inserted into the membrane of the endoplasmic reticulum, and all span the membrane. The size of the cytoplasmic portion of DR alpha and DR beta is about 500- 1000 daltons, whereas that of the DR gamma chain is about 3000 daltons. Oligomeric assembly of DR alpha, DR beta and DR gamma chains occurs shortly after their synthesis in the endoplasmic reticulum. DR gamma chains are synthesized in excess of DR alpha and DR beta chains, and hence in the endoplasmic reticulum they are found either in a complex with DR alpha and DR beta or in a free form. Free DR gamma chains remain in the endoplasmic reticulum, whereas DR gamma chains present in the oligomeric complex with DR alpha and DR beta undergo intracellular transport. Their molecular weight increases during transport, probably because of the addition of complex sugars in the Golgi complex. This is followed by the detachment of DR gamma chains from the oligomeric complex and the appearance of DR alpha and DR beta chains on the cell surface. Whether any DR gamma chains appear on the cell surface is uncertain.