Two types of endogenous ecotropic murine leukemia viruses (MuLVs), termed AKV-and Cas-E-type MuLVsr gene, infectious MuLVs were isolated in association with three of six Cas-E-type endogenous MuLV loci. The isolated viruses showed a weak syncytium-forming activity for XC cells, an interfering property of ecotropic MuLV, and a slight antigenic variation. Two genomic DNAs containing endogenous Cas-E-type MuLV were cloned and partially sequenced. All of the Cas-E-type endogenous MuLVs were closely related, hybrid-type viruses with an ecotropic env gene and a xenotropic long terminal repeat. Duplications and a deletion were found in a restricted region of the hypervariable proline-rich region of Env glycoprotein.Multiple DNA copies of the murine leukemia virus (MuLV) genome, called endogenous MuLV, are present in the chromosomal DNA of Mus musculus (M. m.) mice. MuLVs are largely classified into four groups (ecotropic, xenotropic, amphotropic, and polytropic [or dualtropic] viruses) on the bases of the host range and the interfering properties that are mainly determined by the receptor-binding specificity of the viral envelope (Env) glycoprotein. Ecotropic MuLVs infect mouse cells expressing the receptor for ecotropic MuLVs, mCAT-1 (1). There are two types of endogenous ecotropic MuLVs with a slight sequence divergence. AKV-type endogenous MuLV is carried by many laboratory strains of mice and has been well characterized. Castaneus ecotropic (Cas-E-type) MuLVs are not carried by common laboratory mice and therefore have not been characterized in detail. They are carried by geographically separated M. m. subspecies; AKV-type MuLVs were found in M. m. musculus mice populating the northern part of China, Korea, and Japan, and Cas-E-type MuLVs were found in M. m. castaneus mice populating southern Asia (from Pakistan to Japan) (23, 34). They appear to have diverged along with the subspeciation of M. musculus, probably separated by more than 10 5 to 10 6 years (23). Infectious Cas-E-type MuLVs were isolated from mice from limited areas of California in the United States (6, 9, 15; for a review, see reference 8). Interestingly, unlike AKV-type MuLVs, infectious Cas-E-type MuLVs were reported to be transmitted through sex and milk but not in the germ line in this wild mouse population (10, 11). However, it is not clear by these studies whether endogenous Cas-E-type MuLVs can produce infectious viruses and whether the previously isolated infectious Cas-E-type MuLVs are directly related to endogenous MuLVs.The Fv-4 r gene is a truncated Cas-E-type endogenous MuLV (19,22,41), which is highly homologous in the env gene to infectious Cas-Br-E MuLV (46)