The regulation of HIV-1 transcription is a complex process influenced by host cellular factors and signaling pathways. In this study, we identified Yes-associated protein (YAP) as a novel negative regulator of HIV-1 long terminal repeat (LTR) activity, with the TEAD-binding domain (TBD) of YAP playing a pivotal role in this repression. Mechanistically, YAP suppresses HIV-1 LTR-driven transcription by interacting with the NF-κB binding motif within the LTR, mediated by its downstream TEAD proteins. Notably, our findings reveal that the crosstalk between YAP-TEAD and peroxisome proliferator-activated receptor (PPAR) further modulates HIV-1 LTR transcription, with PPARγ inhibitors reversing YAP-mediated suppression. We show that both YAP and PPAR contribute to the repression of HIV-1 transcription through distinct but complementary mechanisms. This study highlights a previously unrecognized regulatory pathway involving YAP-TEAD-PPAR signaling that suppresses HIV-1 transcription and provides potential new targets for therapeutic intervention.