The pope spoke at the end of his weekly general audience December 6, the same day that Trump was expected to announce his decision to move the US embassy to Jerusalem from Tel Aviv, fulfilling a promise he made during his presidential campaign.
Pope Francis said he could not "keep silent about my deep concern" for Jerusalem and urged respect for "the status quo of the city in accordance with the relevant resolutions of the United Nations".
The position of the United Nations on the Jerusalem issue is that East Jerusalem is occupied Palestinian territory, and that the city should eventually become the capital of the two states of Israel and Palestine.
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(Vatican Radio) Ahead of an expected decision by the U.S President on Wednesday to recognize Jerusalem as the new Israeli capital, a top Vatican official said the move will add "fuel to the fire" of conflict and instability across the Middle East.
While West Jerusalem is the seat of Israel's government, Palestinians view East Jerusalem as the capital of a future Palestinian state.
The pope is effectively telling the president, without naming him, not to proceed down this road, warning that it can lead to even more conflict in the Holy Land and the Middle East.
The pope spoke by telephone to Palestinian President Mahmoud Abbas about the crisis on Tuesday. He told thousands of pilgrims from all continents of his "deep concern" at the proposed change in the US policy toward Jerusalem which is considered sacred to Jews, Christians and Muslims. The Vatican said the call with Francis was made at Abbas' initiative.
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Describing the city as "holy for Jews, Christians and Muslims, who venerate the holy sites of their respective religions", the Pope said Jerusalem enjoys a unique status that should be preserved.
Francis pointed to the meeting with representatives of the various religious communities in the nation as a crucial step toward peace and encouraged further interreligious dialogue between Catholics and Buddhists to fight evil and promote peace.
According to the 1993 Israel-Palestinian peace accords, the final status of Jerusalem will be discussed in the late stages of the talks.
The Pope called for dialogue that would respect the rights and interests of all of Jerusalem's inhabitants in a separate statement made earlier on Wednesday.
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We got the job done but obviously we have a lot of work to do before we can affect that level. "We're looking forward to it". He said: "You have to score a second goal to make them doubt more".
"I wanted, also in this case, to express the closeness of Christ and of the Church to a people that has suffered due to conflict and repression, and which now is slowly walking toward a new condition of freedom and peace", he said.