Recent research has shown that Tooley was born in late 1582 or early 1583; his birth name was not Tooley but Wilkinson. He has been associated with the "Nick" in the surviving "plot" of The Seven Deadly Sins, dated c. 1591. The association, if accurate, indicates that he began as a boy player. He was apprenticed to Richard Burbage, and may have followed that actor to the Lord Chamberlain's Men when that company re-formed in 1594. Tooley is mentioned in a letter of Joan Alleyn, Edward Alleyn's wife, in 1603, and he received a 20-shilling bequest in Augustine Phillips's 1605 will. He became a sharer in the King's Men in 1605, replacing the short-lived Samuel Crosse.
Little is known about Tooley's specific roles for the company. He appears in speech prefixes in the First Folio text of The Taming of the Shrew, and in the cast lists for Ben Jonson's The Alchemist (1610), Sejanus (the 1610 revival), and Catiline (1611). In the revival of John Webster's The Duchess of Malfi staged shortly before his death, he played Forobosco and a madman. In the 25 cast lists added to the second Beaumont and Fletcher folio of 1679, Tooley is mentioned in 14, those for:
His total is lower than those of company stars like Joseph Taylor and John Lowin, but greater than those for most of the King's men's supporting players; Tooley was clearly a significant member of the company.
(The manuscript of the 1619 play Sir John van Olden Barnavelt indiactes that Barnavelt's wife was played by an actor called "Nick." Early critics tended to identify this "Nick" as Tooley, though it may have been Nick Underwood or another unknown player. The question impinges upon the long-debated issue of whether women's roles in English Renaissance drama were filled exclusively by boys, or sometimes by adult actors like Tooley.)
Tooley witnessed Richard Burbage's will in 1619. In his own will of June 3, 1623, Tooley names Henry Condell and Cuthbert Burbage as his executors and residuary legatees. The bequests in Tooley's will are interesting for the light they throw on the actors of the King's Men, and the close relationship Tooley shared with the Burbage family. Those bequests include: