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Knowledge regarding tissue\u2010specific estrogen signaling is important to aid the development of new tissue\u2010specific treatments. We hypothesized that the posttranslational modification phosphorylation in estrogen receptor alpha (ER\u03b1) may modulate ER\u03b1 activity in a tissue\u2010dependent manner. Phosphorylation of site S122 in ER\u03b1 has been shown in vitro to affect ER\u03b1 activity, but the tissue\u2010specific role in vivo is unknown. We herein developed and phenotyped a novel mouse model with a point mutation at the phosphorylation site 122 in ER\u03b1 (S122A). Female S122A mice had increased fat mass and serum insulin levels but unchanged serum sex steroid levels, uterus weight, bone mass, thymus weight, and lymphocyte maturation compared to WT mice. In conclusion, phosphorylation site S122 in ER\u03b1 has a tissue\u2010dependent role with an impact specifically on fat mass in female mice. 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