Background Job satisfaction is an important measure of cognitive as well as a behavioral component of employees’ feelings of comfort in their particular work. It is also the outlook and spirit of certain professionals towards their work. Nurses’ job satisfaction is a significant predictor of staff retention. Therefore, this study aimed to assess Nurse Practitioners’ Job Satisfaction and Associated Factors among Nurses Working at the University of Gondar Specialized Hospital Northwest Ethiopia.Methods A cross-sectional study design was employed from May 15–20/2019. A total of 416 nurses participated in the study. A simple random sampling technique was used to select each study participant. A pretested structured questionnaire was used to collect the data. A Minnesota satisfaction measurement tool was used to measure nurse practitioners’ job satisfaction. A binary logistic regression analysis was used. Variables having p-value < 0.05 in a multivariable analysis model were considered as statistically significant and independently associated with the outcome variable.Result A total of 416 study participants were used in the study with a response rate of 97.6%. In this study, 204(50.2%) of nurse practitioners were dissatisfied with their job. Motivational factors such as professional advancement (AOR = 2.737, 95% CI; 1.109, 6.751), work itself (AOR = 4.250, 95%CI; 1.645, 10.983), recognition at work (AOR = 11.869, 95% CI; 4.238, 33.241), growth at work (AOR = 6.272, 95% CI; 2.278,17.264), and hygienic factors; organizational policy (AOR = 5.623, 95% CI; 2.131, 14.836), relationship with friends (AOR = 6.112,95% CI; 2.393, 15.612), work-security (AOR = 6.562, 95% CI; 95% CI; 2.374,18.136), relationship with supervisor(AOR = 5.127, 95% CI; 1.796, 14.641), payment (AOR = 5.889, 95% CI; 1.989, 17.438), and working conditions AOR = 3.806, 95% CI; 1.195, 12.123) were the factors associated with nurses’ job dissatisfactionConclusions In this study, one of every two nurses experienced job dissatisfaction. Motivational factors such as professional advancement, work itself, recognition at work, growth at work, organizational policy, relationship with friends, work security, relationship with supervisor, payment, and working conditions were the factors associated with nurses’ job dissatisfaction.