This study investigated the relationship between the Σ3 boundaries, dislocation slip, and plasticity in pure nickel wires after grain boundary (GB) modification. Both quasi in situ tensile tests and simulations were employed. During plastic deformation, twins surrounded by Σ3 boundaries may exhibit a good deformation coordination. With an increase in strain, the slip systems corresponding to the maximum Schmid factor and the actual activated slip systems remain unchanged. Even sub-grains can maintain the dominant slip system of their origin matrix grains. Slip systems with slip planes (111) and (1−1−1) are the most active. Moreover, random boundaries have strong hindering effects on dislocations, and the nearby stress accumulates continuously with an increase in strain. In contrast, Σ3 boundaries demonstrate weak blocking effects and can release the nearby stress due to their unique interfacial structures, which is favorable for improving plasticity. They are more penetrable for dislocations or may react with the piled dislocations. In addition, some Σ3 boundaries can improve their geometrical compatibility factor with an increase in the strain, which enhances the deformation coordination of the grains. The research results provide a better understanding of the plasticizing mechanism for face-centered cubic (fcc) materials after grain boundary modification.