Disclosed are an amphiphysin-I mutant having anti-senescence activity and the use thereof. More particularly, disclosed are an amphiphysin-I mutant (AMPH-I) wherein valine (V), which is the 392nd amino acid in the amino acid sequence of amphiphysin-I (AMPH-I) represented by SEQ ID NO: 1, is substituted with glycine (G), a composition for suppressing aging and cellular senescence containing the amphiphysin-I mutant as an active ingredient, a pharmaceutical composition for preventing or treating senescence or a senescence-associated disease, a method for suppressing aging and cellular senescence, and a method for screening an inhibitor for aging and cellular senescence. The amphiphysin-I mutant is capable of suppressing both promotion of aging and cellular senescence and reduction of endocytosis caused by suppression of expression of βPIX (PAK1-interacting exchange factor beta), of preventing cleavage of the amphiphysin-I protein caused by calpain, a protease involved in aging and cellular senescence caused by suppressed βPIX expression, and of suppressing the expression of aging and cellular senescence indicators. Thus, the amphiphysin-I mutant is effectively used as a novel therapeutic agent for senescence or senescence-associated diseases.