The properties of recombinant p66/p51 human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) containing C-terminal truncations in its p66 polypeptide were evaluated. Deletion end points partly or completely removed -helix E` of the RNase H domain (p668/p51 and p6616/p51, respectively), while mutant p6623/p51 lacked E` and the 5`-E` connecting loop. Although dimerization and DNA polymerase properties of all mutants were not significantly different from those of the parental enzyme, p6616/p51 and p6623/p51 RT lacked ribonuclease H (RNase H) activity. In contrast, RT mutant p668/p51 retained endonuclease activity but lacked the directional processing feature of the parental enzyme. Despite retaining full endoribonuclease function, p668/p51 RT barely supported transfer of nascent(-)-strand DNA between RNA templates representing the 5` and 3` ends of retroviral genome, shedding light on the requirement for the endonuclease and directional processing functions of the RNase H domain during replication.

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