This dissertation establishes a new timeframe for Indigenous participation in world-system expansion in southern Ontario, Canada ca. AD 1550–1650, by investigating how and when people living in three different Indigenous Nations in southern Ontario engaged with the expanding European world-system. This will be accomplished by: 1- Establishing an absolute timeframe for the initiation and development of Indigenous-European interaction through Bayesian chronological modeling of high-precision AMS radiocarbon dates from archaeological sites occupied by the Wendat, Tionontate, and Attiwandaron Nations in southern Ontario ca. AD 1550–1650; and 2- investigating both the local and regional spatial distribution of trade goods of European manufacture in these Nations through diversity and evenness analysis of assemblages from Iroquoian sites in southern Ontario AD 1550–1650. Results support the hypothesis that Indigenous people living within different site relocation clusters and different Nations engaged in world-system expansion at different times and in different ways. In particular, Tionontate and Wendat groups at the Western and eastern ends of Lake Ontario, respectively, were involved in the fur trade by the mid-sixteenth century, but with evidence suggesting that Pretty-Mad River Tionontate played a supplier role, while Trent Valley Wendat may have acted as intermediaries. Broadly, this research contributes to debates surrounding the historical development of world-systems, including the degree to which Indigenous agency and differing concepts of value impact the tempo of social change and continuity during the process of colonialism.