Miss Spider, as a giant, speaks with a French accent and wears a beret. She is soft-spoken, brave, tough, feisty, kind, strong and confident. She is like a mother figure to James because he saved her from being killed by his cruelaunts. She initially preferred to be alone, but later warms up. Her love interest is Mr. Centipede, who often tries her patience, but she soon falls in love with him.
She is first seen as an ordinary spider in James' window. He tells her it is a bad idea to build her web there because his aunts might see her and try to kill her. He introduces himself to her and expresses his desire to go to New York and shows her a special candle his parents gave him.
The next morning, Spiker and Sponge spot her, but James saves her, carries her outside, and tells her to leave as fast as she can. That night, James climbs inside the peach, having been transformed, and Miss Spider, who has been magically transformed into a giant along with the other insects, assures him that no one will hurt him, although he does not recognize her.
During their journey to New York City, she aids in dangerous situations, such as fending off a robot shark, and while doing so, she gets James to realize that she is the same spider whose life he had saved. She later starts a romantic relationship with Centipede after he saves her and James from some Skeleton pirates. Although she and the others are separated from James following an attack from the Rhino, they reunite with him and aid him in the capture of Spiker and Sponge, who had journeyed to New York to get James and the peach back. Miss Spider binds James' aunts up in her webbing, and they are taken away. She then lives in Central Park with James and the others, and becomes the owner of "Spider Club."