During meiosis, homologous chromosomes recombine and become closely apposed along their lengths within the synaptonemal complex (SC). In part because Spo11 is required both to make the double-strand breaks (DSBs) that initiate recombination and to promote normal SC formation in many organisms, it is clear that these two processes are intimately coupled. The molecular nature of this linkage is not well understood, but it has been proposed that SC formation initiates locally at the sites of ongoing recombination and in particular at the subset of sites that will eventually give rise to crossovers. To test this hypothesis, we examined further the relationship between DSBs and SC formation in Saccharomyces cerevisiae. SCs were monitored in a series of spo11 missense mutants with varying DSB frequencies. Alleles that blocked DSB formation gave SC phenotypes indistinguishable from a deletion mutant, and partial loss-of-function mutations with progressively more severe DSB defects caused corresponding defects in SC formation. These results strongly correlate SC formation with Spo11 catalytic activity per se. Numbers of Zip3 complexes on chromosomes, thought to represent the sites of SC initiation, also declined when Spo11 activity decreased, but in a markedly nonlinear fashion: hypomorphic spo11 alleles caused larger defects in DSB formation than in Zip3 complex formation. This nonlinear response of Zip3 closely paralleled the response of crossover recombination products. The quantitative relationship between Zip3 foci, SC formation, and crossing over strongly implicates crossover-designated recombination intermediates as the sites of SC initiation.T wo prominent features of meiotic prophase are homologous recombination and formation of the synaptonemal complex (SC). Recombination establishes physical connections between homologous chromosomes in the form of crossovers, which help orient chromosomes properly on the meiosis I spindle. The widely conserved SC connects homologous chromosomes along their lengths during a portion of meiotic prophase and consists of two lateral elements, each formed along the axis connecting a pair of sister chromatids, and a central element linking the lateral elements together (reviewed in refs. 1 and 2).Chromosome synapsis is tightly coupled to homologous recombination in many organisms. The DNA double-strand breaks (DSBs) that initiate recombination occur before synapsis begins in budding yeast (3), and cytological features (e.g., histone H2AX phosphorylation and formation of strand exchange protein complexes) indicate that the same is true in other fungi, higher plants, and mammals (4-8). DSBs are created by the Spo11 protein (9, 10), and spo11 null mutants in many organisms are defective for SC formation (11-16). In some cases, exogenous DSBs can at least partially substitute for Spo11 in SC formation (15)(16)(17). Moreover, mutations that block processing of DSBs once they are formed (e.g., rad50S, dmc1) also cause defects in SC formation (18)(19)(20). These results place Spo11 action...