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The accusation, which follows a US military buildup in the Gulf, came on the eve of emergency Arab and Gulf summits called by Saudi Arabia to discuss the standoff.
Bolton said that additional US forces in the region were sent as a "deterrent" and that Washington's response will be prudent.
The four ships, including two Saudi tankers, were attacked by "naval mines almost certainly from Iran", Bolton told a press conference in the UAE capital. The UAE has not yet blamed anyone for the sabotage of the four vessels near Fujairah, a major bunkering hub just outside the Strait of Hormuz.
"There's no doubt in anybody's mind in Washington who's responsible for this," Bolton said in a clear reference to Iran.
US experts are part of a five-nation team that is investigating the May 12 attacks that damaged the four vessels in the Sea of Oman off the UAE emirate of Fujairah.
Two days later Yemen's Houthi rebels hit a strategic diversionary pipeline in Saudi Arabia with two drones.
The east-west pipeline, which has the capacity to carry some five million barrels per day from the oilfields of the kingdom's Gulf coast to the Red Sea, was shut for two days as a result of the attack.
Bolton said that there has also been "an unsuccessful attack on the Saudi port of Yanbu a couple of days before the attack on tankers".
Yanbu is Saudi Arabia's largest oil terminal on the Red Sea and is home to oil refineries and export facilities.
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