An early play in five acts first performed 1594-95 and published in the First Folio of 1623 from a transcript of a promptbook, probably of a shortened version. It is a pastoral story about two young friends who travel to Milan and are educated in courtly behaviour.

The main source of the play's plot is a translation of a long Spanish prose romance entitled Los siete libros de la Diana (1559?; The Seven Books of the Diana) by Jorge de Montemayor. Shakespeare is thought to have adapted the relationship of the two gentlemen of the title and the ending of the play for Thomas Elyot's The Boke Named the Governour (1531).

Two youths, Valentine and Proteus, make their way from Verona to Milan. They bid a tearful farewell on a street in Verona. Valentine's father is sending Valentine to take a position in the Duke of Milan's court, to improve himself and to all allow him to venturing out to see the world, Proteus meanwhile stays home in Verona, tied by his love for Julia. After Valentine departs, his servant, Speed, enters. Proteus inquires whether Speed delivered a letter to Julia, to which Speed states that he did. Julia, meanwhile, asks her maid, Lucetta, with which man she should fall in love, and Lucetta recommends Proteus. Lucetta admits that she has a letter for Julia from Proteus. After much bickering, Julia tears up the letter, only to regret this act an instant later.

Antonio decides to send Proteus, his son, to the Duke's court in Milan, a decision about which neither Proteus nor Julia is particularly happy. They exchange rings and promises to keep loving each other. Meanwhile, Valentine has fallen in love with the Duke's feisty daughter, Silvia. When Proteus arrives at court, he too falls in love with Silvia, and vows to do anything he can to win her away from Valentine. When Valentine confesses that he and Silvia plan to elope, Proteus notifies the Duke of their plans, gaining favour for himself and effecting Valentine's banishment from court. Back in Verona, Julia has hatched a plan to disguise herself as a man so that she can journey to Milan to be reunited with Proteus. Upon arriving at court, she witnesses Proteus and Thurio wooing Silvia.

The banished Valentine, is apprehended whilst travelling to Mantua, by a group of outlaws. The outlaws, all of whom are also banished gentlemen, demand that Valentine becomes their king. Since they threaten to kill him if he refuses, Valentine accepts.

Julia, ironically now in service as a page to Proteus, becomes an intermediary between Proteus and Silvia. Whilst on a charge of delivering a ring from Proteus to Silvia, on his behalf , Julia, (who is disguised as the page Sebastian), meets Silvia, keeping her identity hidden. Silvia, on the other hand, still longs for Valentine, and cares nothing for Proteus or Thurio. Silvia calls on her friend Sir Eglamour to help her escape her father's oppressive will (he wants her to marry Thurio) and to find Valentine. However, while travelling through the forest, a band of outlaws overwhelms Eglamour and herself. Eglamour runs away, leaving Silvia to fend for herself against the outlaws. By this time, the Duke, Proteus, and Thurio, with Sebastian/Julia in tow, have organized a search party for Silvia.

Managing to find the outlaws before they have managed to carry Silvia back to their encampment, Proteus wrests Silvia away from them. Valentine watches this interaction unseen. Proteus demands that Silvia give him some sign of her favour for freeing her, but she refuses. He tries to rape her for her resistance, but Valentine jumps out and stops him. Proteus immediately apologizes, and Valentine offers to give him Silvia as a token of their friendship. At this moment, Sebastian faints and his true identity becomes clear. Proteus decides that he really loves Julia better than Silvia, and takes her instead. The Duke and Thurio then arrive upon the scene, but Thurio backs off his claim to Silvia when challenged by Valentine. The Duke realizes that Thurio is a thug and says that Valentine is far nobler and can marry Silvia. Valentine asks for clemency for the outlaws, and suggests that his marriage to Silvia and Proteus' marriage to Julia should take place on the same day.

The Two Gentlemen of Verona is a complex and not entirely successful play that shows Shakespeare in the process of developing a new type of high comedy that he would come to master in his middle comedies, especially As You Like It and Twelfth Night.