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., Py-1 (stage 1), Py-2 (stage 2), and Py-WR (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. Py-1 and Py-2 have delta S-34(Sigma S) = 8.28-15.97 parts per thousand (avg. 12.88 parts per thousand) and 6.92-8.70 parts per thousand (avg. 7.67 parts per thousand), respectively, indicating that the sulfur in Py-1 was metamorphic fluid sourced, while that of Py-2 may have a mixed metamorphic fluid and wallrock source (0.84-3.27 parts per thousand; avg. 2.31 parts per thousand). For Py-1, its contents of Au, As, Ag, Bi, Co, Cu, Mn, Ni, Pb, Sb, Tl are the lowest. Py-2 has significantly higher Au-As-Ag, slightly higher Co-Cu-Ni-Sb-Tl, but lower Bi-Mn-Pb contents than those in Py-WR. Considering also the sulfur isotope features, we considered that Py-1 was primarily originated from the initial ore-forming fluid, and Py-2 was derived from both the ore fluid and Py-WR, with the former being more important and represents the source of gold. Based on that Py-2 was formed by metasomatism on the Py-WR 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.