US Deputy Secretary of State William Burns and Deputy Israeli Foreign Minister Danny Ayalon held talks on Thursday focusing on Iran's "quest to develop nuclear weapons," the foreign ministry said.
They discussed "Iran's continued quest to develop nuclear weapons, which the United States and Israel are both determined to prevent, the destabilising role which it plays in the region and its promotion of international terrorism," according to a statement.
Western nations and Israel have accused Iran of seeking to develop nuclear weapons under the guise of a civilian uranium enrichment programme, charges adamantly denied by Tehran.
The meeting at the Israeli foreign ministry, was part of the semi-annual Strategic Dialogue between Israel and the United States.
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The two delegations also agreed that the ongoing bloodshed in Syria, "assisted by Iran and Hezbollah, is a source of major humanitarian concern" and "could also lead to severe consequences for the entire region."
The dialogue "highlights the strong alliance and deep ties" between the two countries, the statement said.
On Wednesday, Burns met with Palestinian president Mahmud Abbas to discuss the stalled peace process with the Israelis.
Direct peace negotiations have been on hold since late September 2010 following an intractable dispute over settlements.
The Palestinians are demanding that Israel halt construction on land they want for a future state and accept a framework for talks on borders.
US Secretary of State Hillary Clinton is due in Jerusalem for talks with Israeli officials on Monday and Tuesday, her first visit for almost two years.