Near the end of a frantic Sunday that saw the lead change hands 19 times, Rose bogeyed the 16th hole to fall into a tie with Phil Mickelson and Hunter Mahan at 1-over.
Rose would go on to par his final two holes and could not watch from the clubhouse to see if his lead would hold up.
Mickelson and Mahan, meanwhile, both stumbled down the stretch to hand the Englishman the title.
Their troubles both began at the 15th, where Mickelson posted a bogey and Mahan recorded a double-bogey to fall behind. As Mahan continued to drop shots at the 17th and 18th to fall well off the pace, Mickelson remained a shot behind with back-to-back pars at the 16th and 17th, then failed to chip in for a tying birdie at the 18th en route to his record-setting sixth runner-up finish at the U.S. Open.
This year, Rose will try to become the first player since Curtis Strange in 1988 and 1989 to win consecutive U.S. Opens.
Mickelson is looking to finally break through and complete the career grand slam when he returns to Pinehurst for the third time. In his two previous U.S. Opens at Pinehurst, Mickelson finished in second place in 1999 and in 33rd in 2005.
Payne Stewart won the first time Pinehurst hosted the championship in 1999 and Michael Campbell captured the title in 2005.
Three-time champion Tiger Woods remains sidelined because of back surgery and will miss his first U.S. Open since 2011. He had finished tied for third in 1999 and second in 2005 in the previous two U.S. Opens played at Pinehurst.
ESPN will have early television coverage on Thursday and Friday before handing off to NBC for the afternoon. ESPN will then take back over for the evening. NBC will have full coverage of the final two rounds.
The PGA Tour will travel to Connecticut next week for the Travelers Championship, where Ken Duke will defend his title. The European Tour returns next week with the Irish Open, which Paul Casey won in 2013.