Use a BO rule for that, something like this:
IF BO IS NEW AND BO.Flag='Yes' then
FIND BO WHERE (BO.ID<>ThisBO.ID AND BO.Flag<>'No')
OtherBO.Flag='No'
If the new BO instance you create thus has the flag set to 'Yes', this rule would find all other instances of the BO that have a 'Yes' flag and set it to 'No'.
You could also do a more general rule which is possibly better, something like this:
IF BO.Flag WAS CHANGED TO 'Yes' AND EXISTS BO WHERE(BO.ID<>ThisBO.ID AND BO.Flag='Yes') then
FIND BO WHERE(BO.ID<>ThisBO.ID AND BO.Flag='Yes')
OtherBO.Flag='No'
This would work in more scenarios e.g if the user sets the new BO flag as 'Yes' it applies but it also applies if the user change the flag later on in some other operation i.e when the BO instance is not new but is just changed. This rule would ensure you always just have 1 BO instance with a 'Yes' flag in the system.