The Kumishi area is located in the eastern part of the South Tianshan Orogen, which hosts several gold deposits and has substantial gold discovery potential. The timing of gold mineralization at Kumishi, however, has been poorly constrained owing to the absence of suitable dating minerals. Hydrothermal activity at Wuzunbulake is divided into the pre-ore stage 1 pyrite-quartz, syn-ore stage 2 quartz(-sulfide) and post-ore stage 3 quartz-calcite alteration/mineralization. Three types of pyrite have been recognized, i.e., Py1 (stage 1), Py2 (stage 2), and PyWR (from wallrock). Our in-situ RbSr dating on stage 2 sericite yielded an isochron age of 351.0 ± 17.4 Ma, indicating Early Carboniferous gold mineralization. Py1 and Py2 have δ34SΣS = 8.28–15.97 ‰ (avg. 12.88 ‰) and 6.92–8.70 ‰ (avg. 7.67 ‰), respectively, indicating that the sulfur in Py1 was metamorphic fluid sourced, while that of Py2 may have a mixed metamorphic fluid and wallrock source (0.84–3.27 ‰; avg. 2.31 ‰). For Py1, its contents of Au, As, Ag, Bi, Co, Cu, Mn, Ni, Pb, Sb, Tl are the lowest. Py2 has significantly higher Au-As-Ag, slightly higher Co-Cu-Ni-Sb-Tl, but lower Bi-Mn-Pb contents than those in PyWR. Considering also the sulfur isotope features, we considered that Py1 was primarily originated from the initial ore-forming fluid, and Py2 was derived from both the ore fluid and PyWR, with the former being more important and represents the source of gold. Based on that Py2 was formed by metasomatism on the PyWR margin and the element spatial coupling characteristics shown in EPMA geochemical maps, we inferred that the Au enrichment and precipitation are associated with fluid-rock reactions. The initial ore fluid is likely featured by the enrichments in Au, As, Ag, Co, Cu, Ni, Sb, Tl, and depletions in Bi, Mn, and Pb. The Wuzunbulake is best classified as an orogenic gold deposit based on its tectonic background, wallrock alteration style, and the ore-fluid source and characteristics.