Rodriguez also faced questions about his gambling habits in 2005, when the Daily News reported he attended games at an underground poker club in New York. The All-Star third baseman later acknowledged "it wasn't the right thing to do," and checked in with Major League Baseball before holding a charity poker tournament the following year.
Now the league wants to talk to him again.
"We take this very seriously and have been investigating this matter since the initial allegation," Major League Baseball said Wednesday in a release. "As part of the investigation, the commissioner's office will interview Mr. Rodriguez."
Star Magazine reported last month that several people saw A-Rod playing in games hosted at Hollywood hotels and residences; his publicist then denied that he participated.
Rodriguez's name is not mentioned in any court filings in relation to the games.
MLB said then that the report was the first the commissioner's office had heard about the accusation and baseball officials would look into the matter.
A spokesman for the Yankees declined comment when asked about Rodriguez on Wednesday. A message was left seeking comment from Richard Rubenstein, Rodriguez's publicist.
The 36-year-old Rodriguez had right knee surgery on July 14 but is expected to resume baseball activities on Thursday at the Yankees' facility in Florida. The three-time AL MVP is hitting .295 with 13 home runs and 52 RBIs this season