Nicholson next served as lieutenant governor or governor of the colonial Provinces of Virginia and Maryland. He supported the founding of the College of William and Mary, at Williamsburg, Virginia, and quarreled with Andros after Andros was selected over him as Governor of Virginia. In 1709, he became involved in colonial military actions during Queen Anne's War (1702-1713, War of the Spanish Succession in Europe), leading an aborted expedition against the French in New France (modern Canada). He then led the expedition that successfully captured Port Royal, Acadia (Nova Scotia) on 2 October 1710. Afterward he served as governor of Nova Scotia and Placentia, and was the first royal governor of the colonial Province of South Carolina following a rebellion against its proprietors. He rose to the rank of Lieutenant-General, and died a bachelor in London, England, in 1728.