After this first colony was abandoned in 1586, Raleigh was anxious to make another attempt at colonization in the New World. This time, he planned to seat the colony on the Chesapeake Bay, north of his first attempt on Roanoke Island. To lead this endeavor, White was chosen as governor of the colony, and in July 1587 the expedition anchored off the Outer Banks of North Carolina near Roanoke Island. The intention of this stop was to check on the approximately fifteen men placed there in 1586 by Richard Grenville, who had left them there the previous year after finally returning in July or August with supplies for the first colonists. Finding the colonists gone, Drake having taken them back to England, Grenville left another small contingent to keep an English presence in the region. When the next group of settlers arrived with White as governor, no trace of Grenville's men was to be found.
White's tenure as governor of the Roanoke colony began against his own wishes as well as Raleigh's. As mentioned, their aim was to settle this second colony on the Chesapeake Bay. When White and the new colonists arrived at Roanoke Island, they disembarked to search for Grenville's men; but when they attempted to return to their ships, their pilot, a Portuguese privateer by the name of Fernandez, would not allow them back on board. With little room for maneuver, White returned to the site of the previous colony at the northern end of Roanoke Island and began to build anew. The previous colony, led by Ralph Lane, had been made up mostly of military men who had little desire for or skill in dealing with the indigenous peoples. That attitude resulted in hostilities between the Native Americans and the English, and White's colonists soon began to suffer the hardships of living in an alien and hostile environment with too few supplies or skills. The colonists, among them White's daughter and granddaughter, agreed that White should return to England for supplies and relief. He did so in August of 1587 with hopes of a rapid return. Other forces were at work, however, and his anticipated quick return became an agonizing wait.