Alphatogaviruses, of which Sindbis virus (SV) is the prototype, replicate to high titer in the laboratory both in mosquito cells and in vertebrate cells. By studying the replication of SV in mosquito cells as well as in vertebrate cells, we were able to obtain several viral mutants which have novel phenotypes and have contributed to our basic knowledge of this virus family. These include three host range mutants: SVAP15/21 which replicates normally in mosquito cells but is restricted in vertebrate cells and SVCL35 and SVCL58, which are restricted in mosquito cells but replicate normally in vertebrate cells. As well, two other mutants are described here: SVLM21, which can replicate in methionine-starved mosquito cells and SVMPA, which can replicate in mosquito cells treated with mycophenolic acid or ribavirin. The causal mutations of both SVLM21 and SVMPA are within the sequence encoding the nonstructural protein nsPl; these and other findings have enabled us to associate the capping and methylation of the viral mRNAs with the nsPl protein. Our work serves to emphasize that it is both worthwhile and important to study the replication of arthropod-borne viruses in cells derived from the arthropod host as well as in cells derived from the vertebrate host.