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Strain engineering provides an effective way of tailoring the electronic and optoelectronic properties of semiconductor nanomaterials and nanodevices, giving rise to novel functionalities. Here, we present direct experimental evidence of strain-induced modifications of hole mobility in individual GaAs nanowires, using in situ transmission electron microscopy (TEM). The conductivity of the nanowires varied with applied uniaxial tensile stress, showing an initial decrease of ~5-20% up to a stress of 1~ 2 GPa, subsequently increasing up to the elastic limit of the nanowires. This is attributed to a hole mobility variation due to changes in the valence band structure caused by stress and strain. The corresponding lattice strain in the nanowires was quantified by in situ 4D-scanning TEM (STEM) and showed a complex spatial distribution at all stress levels. Meanwhile, a significant red shift of the band gap induced by the stress and strain was unveiled by monochromated electron energy loss spectroscopy.