Heat shock protein 90 (Hsp90) plays a critical role in maintaining the proteostasis of Plasmodium falciparum, the most lethal malaria parasite. Although PfHsp90 is recognized as a promising antimalarial target, its interactome and the global proteomic remodeling induced by its inhibition remain poorly characterized. Herein, we leveraged global chemoproteomic profiling employing two PfHsp90 inhibitors with distinct scaffolds, geldanamycin and XL888, to investigate proteins and pathways dependent on the chaperone during the P. falciparum asexual blood stage. This study revealed 133 hits with significantly decreased abundances in response to PfHsp90 inhibition with both compounds. A subset of these hits demonstrated conservation as Hsp90 interactors in yeast and human model organisms, while a large majority of the hits lacked predicted orthologs. Bioinformatics analyses of the hits yielded strong enrichment in DNA replication and associated processes and this link was confirmed in phenotypic studies demonstrating that PfHsp90 inhibition reduces the total P. falciparum DNA content. To specifically assess nascent DNA synthesis, we utilized a 7-deaza-7-ethynyl-2’-deoxyadenosine (EdA) incorporation assay, which demonstrated impairment of active DNA replication following PfHsp90 inhibitor treatment. We further show that the co-treatment of parasites with PfHsp90 and DNA replication inhibitors produces synergistic interactions, highlighting the therapeutic potential of the discovered link. Together these findings reveal over a hundred putative Hsp90-dependent proteins as possible interactors in the pathogenic parasites, where the functional dependence of P. falciparum DNA replication on PfHsp90 was validated. These efforts expand our understanding of PfHsp90 function and uncover the potential for dual targeting of these pathways in the fight against malaria.