Human immunodeficiency virus type 1 (HIV-1) recruits diverse cellular factors into viral particles during its morphogenesis, which apparently play roles in modulating its infectivity. The article by Mouree et al. evaluated that a key glycolytic protein, pyruvate kinase muscle type 2 (PKM2) is incorporated into viral particles. Furthermore, the virion-packaged PKM2 significantly reduces the viral infectivity by affecting the selective packaging of intravirion tRNALys3, which primes the initiation of reverse transcription, along with other nonpriming tRNAs, such as tRNALys1,2 and tRNAAsn, without affecting the cytoplasmic level of these tRNAs. These findings proposed that PKM2 is a vital host factor that negatively affects HIV-1 infectivity by targeting the tRNALys3-mediated initiation of reverse transcription in target cells.