The effect of cooling rate, ranging from 6 to 1 °C/s, on microstructure and mechanical properties in the coarse-grained heat affected zone (CGHAZ) of electroslag welded pearlitic rail steel has been investigated by using confocal scanning laser microcopy (CSLM) and Gleeble 3500 thermo-mechanical simulator. During heating, the formed austenite was inhomogeneous with fractions of untransformed ferrite, which has influenced the pearlite transformation during cooling by providing additional nucleation sites to pearlite. During cooling, at 6 °C/s, the microstructure was composed of martensite and bainite with little pearlite. From 4 to 1 °C/s, microstructures were completely pearlite. Lowering the cooling rate of the CGHAZ from 4 to 1 °C/s increased the pearlite start temperature and reduced the pearlite growth rate. Meanwhile, this increase in pearlite start temperature enlarged the pearlite interlamellar spacing. Alternatively, increasing pearlite interlamellar spacing in the CGHAZ by lowering the cooling rate from 6 to 1 °C/s reduced the hardness and tensile strength, whereas toughness was found unaffected by the pearlite interlamellar spacing. It has been found that a cooling rate of 4 °C/s leads to the formation of pearlite with fine interlamellar spacing of 117 nm in the CGHAZ of electroslag welded pearlitic rail steel where hardness is 425 HV, tensile strength is 1077 MPa, and toughness is 9.1 J.