Gatlin, running in Lane 8, was timed finishing in 9.92 seconds, followed by Coleman in Lane 4 at 9.94 and Bolt in Lane 3 at 9.95 seconds. A 21-year-old Christian Coleman too went past the Jamaican by a hundredth of a second. However, Bolt was all smiles and gracious in defeat.
Now Bolt is officially retiring, and the sport is left with a huge void for losing its most dazzling star.
Though the relays remain as realistic medal prospects, but as we enter the final two days of the IAAF World Championships there is no escaping the reality that Jamaica's athletes in London have performed below expectations.
The first instance was in 2011 when he faltered with a false start in the 100 metres finals at the World Championship in Daegu, South Korea.
The eight-time Olympic gold medalist finished third in a photo finish to runner-up Christian Coleman, and Gatlin on August 5 at London Stadium, site of the 2012 Olympic Games. The relay heats are set for early Saturday.
So just what did the world make of it? For Bolt, it goes beyond winning.
But for the grand entrance of Bolt into the scene, athletics was nearly being ruined by the several doping scandals that had marred the image of the sport.
Jamaica now leads the Caribbean with three medals -two bronze and one gold, while Cuba, The Bahamas and Trinidad & Tobago have one each.
It was the first time Bolt had been beaten at a major championship since the 4x100m relay at the 2007 worlds in Osaka. For he is just one of a suspicious number of Bolt's Jamaican teammates who, like Gatlin, have returned to competition after testing positive for PEDs.
Unlike Thompson, Bolt decided not to compete in the 200 at these world championships.
Bolt admitted he was disappointed but would never change his idea of retiring because the new life ahead is more inviting.
As it turned out, while Mr Bolt was good enough to take a bronze medal despite being way below the required level of preparedness, Miss Thompson, a double gold medallist at the Rio Olympics previous year, could only manage fifth in London. No regrets, I came out and did my best; win lose or draw. But I dont think anybody was thinking that it is impossible for him to lose, said the Briton. It is the good news story they crave.
But there was no margin for grandstanding in this final. Bolt routinely slowed down before crossing the finish line. "And as he started to decelerate, his face was full of concern as he looked at the scoreboard for the results".
A distraught Bolt was consoled on the ground by his sprint team-mates and he limped out of the stadium, not the way he probably envisaged his send-off going.
Much will be out of Bolt's hands.
This is why it is, indeed, a sad tale for athletics that it was Justin Gatlin, a 35-year-old athlete who has had a drug blemished career that is now the world 100 meters champion.
Michael Johnson, a former world champion, stated that Gatlin was being jeered because of the way the media had presented him to the public. They also added that Bolt prolonged his career for "commercial incentives".