The impending removal of two dams on the Elwha River in Washington State offers a unique opportunity to study ecosystem restoration at a watershed scale. We examine how periphyton and benthic invertebrate assemblages vary across regulated and unregulated sections of the Elwha River and across different habitat types, and establish baseline data for tracking future changes following dam removal. We collected multiple years of data on physical habitat, water chemistry, periphyton, and benthic invertebrates from 52 sites on the Elwha River and a reference section on the Quinault River, a neighboring basin. We found that substrate in regulated river sections was coarser and less heterogeneous in size than in unregulated sections, and summer water temperature and specific conductivity higher. Periphyton biomass was also consistently higher in regulated than unregulated sections. Benthic invertebrate assemblage structure at sites above both dams was distinct from sites between and below the dams, due in large part to dominance of mayfly taxa compared to higher relative abundance of midges and non-insect taxa at downstream sites. Following dam removal, we anticipate that both periphyton and benthic invertebrate abundance and diversity will temporarily decrease between and below dams as a result of sediment released from behind the reservoirs. Over the long-term, increased floodplain heterogeneity and recolonization by anadromous fish will alter benthic invertebrate and periphyton assemblages via increases in niche diversity and inputs of marine-derived nutrients. The extended timeline predicted for Elwha River recovery and the complexities of forecasting ecological response highlights the need for more long-term assessments of dam removal and river restoration practices.