Activation of transcription factor NF-κB is a hallmark of infection with the gastric pathogen Helicobacter pylori, associated with inflammation and carcinogenesis. Genome-wide RNAi screening revealed numerous host factors involved in H. pylori-, but not IL-1β- and TNF-α-dependent NF-κB regulation. Pathway analysis including CRISPR/Cas9-knockout and recombinant protein technology, immunofluorescence microscopy, immunoblotting, mass spectrometry, and mutant H. pylori strains identified the H. pylori metabolite D-glycero-β-D-manno-heptose 1,7-bisphosphate (βHBP) as a cagPAI type IV secretion system (T4SS)-dependent effector of NF-κB activation in infected cells. Upon pathogen-host cell contact, TIFA forms large complexes (TIFAsomes) including interacting host factors, such as TRAF2. NF-κB activation, TIFA phosphorylation, and TIFAsome formation depend on a functional ALPK1 kinase, highlighting the ALPK1-TIFA axis as a core innate immune pathway. ALPK1-TIFA-mediated NF-κB activation was independent of CagA protein translocation, indicating that CagA translocation and HBP delivery to host cells are distinct features of the pathogen’s T4SS. Display omitted •Genome-wide RNAi screen identifies key innate immune pathway induced by H. pylori•Release of LPS metabolite HBP via H. pylori’s type IV secretion activates NF-κB•The ALPK1-TIFA axis constitutes a core pathogen-specific pattern recognition system Zimmermann et al. identify pathogen and host factors involved in NF-κB activation after infection with type IV secretion-proficient Helicobacter pylori. Central hits are ALPK1 and TIFA. ALPK1 is necessary for phosphorylation-dependent formation of TIFA complexes (TIFAsomes) with TRAF2. This yields HBP-ALPK1-TIFA-TRAF2-NF-κB as the core-regulon of H. pylori-induced innate immune activation.