The Obama administration has expelled two Venezuelan diplomats, US officials say, in retaliation for Venezuela's expulsion of two US military attaches.
Shortly before Chavez died last week, Venezuela expelled two US Air Force attaches in Caracas for alleged espionage.
The Obama administration waited until after Chavez's funeral on Friday to announce any reciprocal action.
Monday's move comes as Venezuela prepares for an April election to choose a new leader.
The US and Venezuela have not had ambassadors posted in each other's capitals since 2010.
Chavez rejected the US nominee at the time, accusing him of making disrespectful remarks about the Venezuelan government.
Washington then revoked the visa of Venezuela's ambassador to the US.
On Saturday, US officials said junior Venezuelan diplomats Orlando Jose Montanez Olivares and Victor Camacaro Mata were ordered to return home.
Montanez, an official at the embassy in Washington, and Camacaro, who served in Venezuela's New York consulate, left the United States on Sunday.
The US officials spoke on condition of anonymity because they were not authorised to speak publicly about the expulsions.
Suggestions of poisoning
Beyond the diplomatic tit-for-tat, Venezuelan officials have accused the US of being responsible for Chavez's cancer and sought to rally anti-US sentiment ahead of an April election for a new leader.
Administration officials declared themselves highly disappointed with Nicolas Maduro, the interim president and Chavez's desired successor, for a news conference he gave last week.
Comparing Chavez to Palestinian leader Yasser Arafat, Maduro suggested that Chavez had been poisoned.
In recent months, as Chavez's health deteriorated, the administration sounded out Maduro in an attempt to improve relations that became badly strained during Chavez's 14 years in power.
Despite some positive feedback from a November telephone call with Roberta Jacobson, the top US diplomat for Latin America, American officials see little possibility of a sudden improvement in relations with Venezuela given its upcoming election.
Maduro is running against opposition leader Henrique Capriles.
Officially, Washington has not taken sides. It has focused its calls on the need for free and fair elections.