Deneke, V. E.; Suwita, J. P.; Wang, H.; Tonai, S.; Lu, Y.; Panser, K.; Schleiffer, A.; Hollis, J. A.; Novatchkova, M.; Duernberger, G.; Stejskal, K.; Krssakova, G.; Blaha, A.; Andresan, A. A. R.; Mirus, M.; Marvanova, H.; Chang, H.-Y.; Noda, T.; Burga, A.; Roitinger, E.; Ikawa, M.; Pauli, A.

Fertilization requires gamete recognition and membrane fusion, yet the molecular basis of this process in vertebrates remains unknown. Here we identify SPARK (sperm protein assembly and receptor-binding key), a conserved multi-protein complex that integrates all known sperm fertilization factors, including TMEM81-IZUMO1-SPACA6 and DCST1/2, together with two newly identified components, TMDD1 and FAM187A. SPARK subunits are mutually dependent for stability in mature sperm, and disruption of any single component causes male sterility in zebrafish and mice. Incubating zebrafish sperm with soluble egg receptor Bouncer partially rescues fertilization of Bouncer-deficient eggs in a SPARK-dependent manner, consistent with egg receptor binding priming the complex for fusion. Thus, we propose SPARK as a conserved molecular machine that couples gamete recognition to membrane fusion.