Nick Easter has been replaced in England's senior squad by uncapped Northampton forward Courtney Lawes after losing his battle with a calf injury.
Easter tore the muscle in his right leg in Harlequins' 26-22 Guinness Premiership victory over Worcester on Saturday and has been ruled out for four weeks.
It is the latest blow to a squad that has been heavily disrupted by injury, but Easter's absence has enabled Lawes to continue his rapid rise up the ranks.
Lawes' promotion from the Saxons to elite player squad is one of 11 straight changes, which include the return of World Cup winners Lewis Moody and Steve Thompson.
A replacement has yet to be named for injured prop Julian White, but Stade Francais number eight James Haskell is back in the fold to complete the 32.
Bath's veteran props Duncan Bell and David Barnes are included to address the front-row crisis, while winger David Strettle, fly-half Andy Goode and Shane Geraghty also return.
There is also a place in the squad for uncapped Leicester second row Richard Blaze.
The 24-year-old's presence has come at the expense of Simon Shaw, who is recovering from a fractured metatarsal and may even struggle to be ready for the concluding Test against New Zealand on November 21.
Saracens lock Steve Borthwick has been confirmed as skipper for the series, which opens against Australia on November 7 and includes a fixture against Argentina a week later.
"The whole squad is looking forward to three very tough games in November," said Johnson.
"Steve has been appointed captain and I'm sure he will continue to do an excellent job.
"We've had to make a number of changes to the senior EPS because of injuries but we have strength in depth.
"Each of the players coming in has the opportunity to push themselves into contention for the opener against Australia."
England's revised senior EPS squad for Investec Challenge Series matches against Australia, Argentina and New Zealand:
Forwards: S Armitage (London Irish), D Barnes (Bath), D Bell (Bath), R Blaze (Leicester), S Borthwick (Saracens, captain), G Chuter (Leicester), J Crane (Leicester), T Croft (Leicester), L Deacon (Leicester), D Hartley (Northampton), J Haskell (Stade Francais), B Kay (Leicester), C Lawes (Northampton), L Moody (Leicester), T Payne (Wasps), S Thompson (Brive), D Wilson (Bath), J Worsley (Wasps).
Backs: M Banahan (Bath), D Care (Harlequins), M Cueto (Sale), B Foden (Northampton), S Geraghty (Northampton), A Goode (Brive), D Hipkiss (Leicester), P Hodgson (London Irish), U Monye (Harlequins), D Strettle (Harlequins), M Tait (Sale), M Tindall (Gloucester), R Wigglesworth (Sale), J Wilkinson (Toulon).
England flanker James Haskell insists he will be pushing as hard as possible to be back from a toe injury in time to make his case for a spot in Eddie Jones' side looking to defend their RBS 6 Nations title next year.