Dataset Information


Second-site changes affect viability of amphotropic/ecotropic chimeric enveloped murine leukemia viruses.

ABSTRACT: Chimeras were previously generated between the ecotropic (Moloney-MuLV) and amphotropic (4070A) SU and TM proteins of murine leukemia virus (MuLV). After passage in D17 cells, three chimeras with junctions in the C terminus of SU (AE5, AE6, and AE7), showed improved kinetics of viral spreading, suggesting that they had adapted. Sequencing of the viruses derived from the D17 cell lines revealed second-site changes within the env gene. Changes were detected in the receptor binding domain, the proline-rich region, the C terminus of SU, and the ectodomain of TM. Second-site changes were subcloned into the parental DNA, singly and in combination, and tested for viability. All viruses had maintained their original cloned mutations and junctions. Reconstruction and passage of AE7 or AE6 virus with single point mutations recovered the additional second-site changes identified in the parental population. The AE5 isolate required changes in the VRA, the VRC, the VRB-hinge region, and the C terminus of SU for efficient infection. Passage of virus, including the parental 4070A, in D17 cells resulted in a predominant G100R mutation within the receptor binding domain. Viruses were subjected to titer determination in three cell types, NIH 3T3, canine D17, and 293T. AE6 viruses with changes in the proline-rich region initially adapted for growth on D17 cells could infect all cell types tested. AE6-based chimeras with additional mutations in the C terminus of SU could infect D17 and 293T cells. Infection of NIH 3T3 cells was dependent on the proline-rich mutation. AE7-based chimeras encoding L538Q and G100R were impaired in infecting NIH 3T3 and 293T cells.


PROVIDER: S-EPMC111611 | BioStudies | 2000-01-01

REPOSITORIES: biostudies

Similar Datasets

2003-01-01 | S-EPMC140616 | BioStudies
2001-01-01 | S-EPMC114680 | BioStudies
1000-01-01 | S-EPMC241488 | BioStudies
2010-01-01 | S-EPMC2998453 | BioStudies
1000-01-01 | S-EPMC241286 | BioStudies
2003-01-01 | S-EPMC152129 | BioStudies
1998-01-01 | S-EPMC124602 | BioStudies
1999-01-01 | S-EPMC112780 | BioStudies
1000-01-01 | S-EPMC4938242 | BioStudies
2011-01-01 | S-EPMC3067788 | BioStudies