The initial settlement was established in the summer of 1585, but a lack of supplies and bad relations with the local Native Americans caused many of its members to return to England with Sir Francis Drake a year later, leaving behind a small detachment. These men had all disappeared by the time a second expedition led by John White, who also served as the colony's governor, arrived in July 1587. White, whose granddaughter Virginia Dare was born there shortly thereafter (making her the first English child born in the New World), left for England in late 1587 to request assistance from the government, but was prevented from returning to Roanoke until August 1590 due to the Anglo-Spanish War. Upon his arrival, the entire colony was missing with only a single clue to indicate what happened to them: the word "CROATOAN" carved into a tree.
For many years, it was widely accepted that the colonists were massacred by local tribes, but no bodies were ever discovered, nor any other archaeological evidence. The most prevalent hypothesis now is that environmental circumstances forced the colonists to take shelter with local tribes, but that is mostly based on oral histories and also lacks conclusive evidence. Some artifacts were discovered in 1998 on Hatteras Island where the Croatan tribe was based, but researchers could not definitively say these were from the Roanoke colonists.
The enterprise was originally financed and organized by Sir Humphrey Gilbert, who drowned in 1583 returning from a voyage to the fishing settlement at St. John's, Newfoundland. Sir Humphrey Gilbert's half-brother Sir Walter Raleigh later gained his brother's charter from the Queen and subsequently executed the details of the charter through his delegates Ralph Lane and Richard Grenville, Raleigh's distant cousin.