GP63 is a 63-kDa glycoprotein that is abundantly expressed on the surface of all Leishmania species and is involved in several steps of promastigote infection of host cells. Leishmania chagasi has at least 18 haploid msp (major surface protease) genes encoding GP63 that are divided into three classes, mspS, mspL or mspC, according to their unique 3' UTR sequences and differential expression. All three msp classes are constitutively transcribed during virulent promastigote growth in vitro, although mspL mRNA is most abundant during logarithmic phase and mspS mRNA predominates in stationary phase. Thus, the steady state levels of the mspL and mspS mRNAs are post-transcriptionally regulated. Using Actinomycin D to arrest transcription, we found that in virulent promastigotes the half-life (t(1/2)) of mspL mRNA is coordinately modulated with growth phase, decreasing from a mean of 84 min during early logarithmic growth to a mean of 17 min at a stage intermediate between logarithmic and stationary phase. However, in attenuated promastigotes, the t(1/2) of mspL RNA remains the same throughout parasite growth. In contrast to mspL RNA, the t(1/2) of mspS and mspC RNA is constant throughout all growth phases of both virulent and attenuated promastigote growth. The presence of the translation inhibitor cycloheximide increases the t(1/2) of mspL RNA 4-6-fold in both virulent and attenuated promastigotes at all growth phases. These results indicate that the t(1/2) of mspL RNA is maintained by at least two distinct mechanisms - one activated during growth to stationary phase and the other dependent on a labile negative regulatory protein factor(s).