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Dysregulated PKA signaling is implicated in diseases including neurodegeneration and cancer. PKA catalytic activity is regulated by two nonredundant regulatory subunits, Type I (RI\u03b1\/RI\u03b2) and Type II (RII\u03b1\/RII\u03b2), whose divergent functions are not fully understood. We generated double-knockout (KO) cell lines of RI\u03b1\/RI\u03b2 and RII\u03b1\/RII\u03b2 subunits and performed multiplexed MS-based proteomic and phosphoproteomic profiling under basal and glucose-perturbed conditions. We found that RI and RII loss drives distinct, and often opposite, remodeling of the cellular proteome and phosphoproteome. While both mutants blunted metabolic flexibility to glycolytic stressors and stimuli, RI and RII KO cells exhibited elevated and depressed glycolytic signaling, respectively. Interestingly, RI KO increased the abundance and kinase activity of the PKA catalytic subunit C\u03b1 isoform, leading to an increase in PKA substrate phosphorylation, whereas RII KO decreased the abundance, kinase activity, and substrate phosphorylation by the catalytic subunit C\u03b2 isoform. Notably, one of the most differentially affected PKA sites between RI and RII KOs maps to Tau, whose hyperphosphorylation is a hallmark of Alzheimer\u2019s disease. Loss of RI increased Tau phosphorylation, which was not only caused by increased PKA catalytic activity, but also a higher binding affinity of Tau to RII subunits on the negatively-charged flexible linker region. Overall, the present study demonstrates that PKA RI and RII subunits play nonredundant roles in modulating PKA activity, metabolic flexibility, and phospho-regulation of key disease-associated substrates such as Tau.<\/jats:p>","DOI":"10.64898\/2026.06.26.734839","type":"posted-content","created":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T13:20:20Z","timestamp":1782739220000},"source":"Crossref","is-referenced-by-count":0,"title":["Selective knockout of PKA regulatory subunits reveal opposite catalytic and metabolic consequences with implications for Alzheimer\u2019s disease"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7744-3630","authenticated-orcid":false,"given":"Leigh-Ana M.","family":"Rossitto","sequence":"first","affiliation":[{"name":"Department of Pharmacology, University of California San Diego, San Diego, CA 92093, USA"},{"name":"Department of Neurosciences, University of California San Diego, San Diego, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tsanwen","family":"Lu","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California San Diego, San Diego, CA 92093, USA"},{"name":"Department of Biochemistry & Molecular Biophysics, University of California San Diego, San Diego, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuliang","family":"Ma","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California San Diego, San Diego, CA 92093, USA"},{"name":"Department of Biochemistry & Molecular Biophysics, University of California San Diego, San Diego, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2986-3515","authenticated-orcid":false,"given":"Pallavi","family":"Kaila Sharma","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California San Diego, San Diego, CA 92093, USA"},{"name":"Department of Biochemistry & Molecular Biophysics, University of California San Diego, San Diego, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valeria","family":"Burghi","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California San Diego, San Diego, CA 92093, USA"},{"name":"Moores Cancer Center, University of California San Diego, San Diego, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4673-4048","authenticated-orcid":false,"given":"Carlos C.","family":"Gonzalez","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California San Diego, San Diego, CA 92093, USA"},{"name":"Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, San Diego, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-8192-814X","authenticated-orcid":false,"given":"Jessica","family":"Bruystens","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California San Diego, San Diego, CA 92093, USA"},{"name":"Department of Biochemistry & Molecular Biophysics, University of California San Diego, San Diego, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4700-2711","authenticated-orcid":false,"given":"Svetlana","family":"Maurya","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California San Diego, San Diego, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8031-9462","authenticated-orcid":false,"given":"Jian","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California San Diego, San Diego, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-6147-3804","authenticated-orcid":false,"given":"Alexis","family":"Lona","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California San Diego, San Diego, CA 92093, USA"},{"name":"Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, San Diego, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9083-7039","authenticated-orcid":false,"given":"Irina","family":"Kufareva","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of California San Diego, San Diego, CA 92093, USA"},{"name":"Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California San Diego, San Diego, CA 92093, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5150-4482","authenticated-orcid":false,"given":"J. 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