South Africa will provide the opposition, with Hartley and Saracens fly-half Owen Farrell co-captaining England in the clash.
Hartley has been in good form for Saints this season, making seven appearances for Chris Boyd's side.
And he has been rewarded by England boss Eddie Jones, keeping Saracens hooker Jamie George on the bench.
But Courtney Lawes will not be involved as he continues to recover from the back problem he picked up during the second half of Saints' win at Bristol Bears on September 29.
On England’s preparation ahead of Saturday, boss Jones said: “We’ve become very well organised in our set piece and have done a lot of good work in Portugal over the last week.
"We have put in a new defence system and our attack looks more organised than it was on the South Africa tour.
“Against South Africa you have got the physical battle up front and then you have to be tactically smart in how you attack against them.
"We need to find ways to gain momentum, then once we find momentum, convert that to points.
“We are really excited to be back at Twickenham.
"It’s been a long time and we can’t wait to play in front of 82,000 fans.”
England starting XV
15 Elliot Daly (Wasps, 21 caps)
14 Jack Nowell (Exeter Chiefs, 26 caps)
13 Henry Slade (Exeter Chiefs, 13 caps)
12 Ben Te’o (Worcester Warriors, 13 caps)
11 Jonny May (Leicester Tigers, 37 caps)
10 Owen Farrell (Saracens, 61 caps) co-captain
9 Ben Youngs (Leicester Tigers, 77 caps)
1 Alec Hepburn (Exeter Chiefs, 2 caps)
2 Dylan Hartley (Saints, 93 caps) co-captain
3 Kyle Sinckler (Harlequins, 13 caps)
4 Maro Itoje (Saracens, 22 caps)
5 George Kruis (Saracens, 25 caps)
6 Brad Shields (Wasps, 2 caps)
7 Tom Curry (Sale Sharks, 4 caps)
8 Mark Wilson (Newcastle Falcons, 4 caps)
16 Jamie George (Saracens, 28 caps)
17 Ben Moon (Exeter Chiefs, uncapped)
18 Harry Williams (Exeter Chiefs, 11 caps)
19 Charlie Ewels (Bath Rugby, 6 caps)
20 Zach Mercer (Bath Rugby, uncapped)
21 Danny Care (Harlequins, 81 caps)
22 George Ford (Leicester Tigers, 47 caps)
23 Manu Tuilagi (Leicester Tigers, 26 caps)