Plasma homocysteine (Hcy) levels may be associated with all-cause mortality risk. However, the results of this association are conflicting and the dose-response relationship between them has not been clearly defined. In this meta-analysis, we conducted a systematic literature search of the PubMed, Embase, Web of Science and Cochrane Library for the relevant articles dated up to February 2017. Pooled relative risks (RRs) and corresponding 95% confidence intervals (CIs) were calculated to evaluate the estimates, and the dose-response relationship was estimated using a restricted cubic spline model. Eleven prospective studies (4,110 deaths among 27,737 individuals) were included. The summary RR of all-cause mortality for the highest Hcy category vs. the lowest Hcy category was 1.80 (95% CI: 1.51, 2.14) with the random effects model. In dose-response meta-analysis, Hcy levels were significantly associated with all-cause mortality risk in a linear fashion (p nonlinearity = 0.255), and the risk of all-cause mortality increased by 33.6% for each 5 µmol/L increase in Hcy levels (RR = 1.336, 95% CI: 1.254-1.422, p < 0.001). Findings from this dose-response meta-analysis suggest that Hcy levels are linearly and positively associated with risk of all-cause mortality.