Sequence analysis of lymphocytic choriomeningitis virus L and S RNAs has revealed evidence of heterogeneity within the termini of the genomic and antigenomic RNAs. The RNAs are missing from 0 to 38 bases, show characteristic patterns of deleted nucleotides at both 5' and 3' termini, and often have a nontemplated base at the terminus. The same deletions, at either the 5' or the 3' terminus of the genomic L and S RNAs, are frequently found in the complementary strand of antigenomic RNA, suggesting that RNAs with deleted termini may be recognized as functional templates for replication. RNAs extracted from virions, or viral nucleocapsids isolated from acutely infected cells, are similar in the nature and extent of terminal heterogeneity that have been observed. This finding brings into question the function of the conserved sequences located at the termini of arenavirus genomic RNAs. Our data suggest that, while replication and packaging of the genomic and antigenomic RNA molecules can occur with terminally deleted molecules, mature transcripts may be derived only from full-length templates containing the conserved terminal sequence.