The 26-year-old has taken the place of Jack Grealish on the left-hand side of the attack, as Gareth Southgate makes three changes to the side that beat Italy 2-1 on Thursday evening.
Harry Kane and Bukayo Saka hold their spots in the final third alongside Maddison, while Jude Bellingham has shaken off an injury scare to start once again in the engine room.
Declan Rice also retains his place after scoring his first England goal in Naples, but Kalvin Phillips - who has played very little for Manchester City this season - drops out for Jordan Henderson.
With Luke Shaw suspended, Ben Chilwell is drafted in at left-back, while Kyle Walker fends off the threat of Kieran Trippier to hold his place on the right-hand side of the backline.
John Stones and Harry Maguire completely a largely unaltered backline alongside number one Jordan Pickford, while Phil Foden withdrew due to appendicitis on Sunday morning.
Meanwhile, Arsenal's Oleksandr Zinchenko and Chelsea's Mykhaylo Mudryk both start for Ukraine, but ex-West Ham United attacker Andriy Yarmolenko is absent due to a thigh injury.
England: Pickford; Walker, Stones, Maguire, Chilwell; Henderson, Rice, Bellingham; Saka, Kane, Maddison
Subs: Ramsdale, Forster, Trippier, Dier, Guehi, Phillips, Gallagher, Toney, Grealish
Ukraine: Trubin; Karavaev, Svatok, Matvienko, Mykolenko; Zinchenko, Stepanenko; Malinovskyi, Sudakov, Mudryk; Yaremchuk
Subs: Lunin, Shevchenko, Sobol, Sarapiy, Sydorchuk, Konoplyanka, Dovbyk, Miroshnychenko, Buyalskyi, Tsygankov, Bondarenko, Pikhalonok