Italy is a predominantly Catholic country that developed historically on the basis of a strong, dominant religion and weak state institutions. Yet, openly clerical parties, direct advocates of the interests of the Catholic Church, have nowadays virtually disappeared and the relevance of the religious cleavage is decreasing, in favour of a more indirect support for these interests, mainly among moderate and conservative forces. Although the overall level of secularisation in Italy has increased, the degree of religiosity of Italian society remains one of the highest of the 27 member-states of the European Union (EU) and polarisation over religious issues in domestic politics remains high, particularly regarding moral values and family matters. In our study we explore the role of religion within the Italian political sphere with regard to the functioning of political representation, by taking into account the sub-national, national and European levels of government. We focus on the attitudes and behaviours of Italian political elites at the EU level. We hypothesise a strong influence of religion on the articulation between national and European politics. Our findings consistently show that the degree of religiosity of the Italian delegation to the European Parliament (EP) is high. However, the impact of such a high degree of religiosity among the members of the EP (MEPs) on their political activities appears less direct than one might predict, while the degree of political secularism is higher among Italian MEPs than among their national or regional counterparts. When we discuss a case study, namely the accession of Turkey to the EU, our data show that the religious attitudes of Italian MEPs play a crucial role in their stance on Turkish accession. The picture that emerges is thus nuanced. Religion significantly impacts on Italian MEPs’ ideological, political and moral attitudes, but plays a smaller role in their activities; while their left-right collocation emerges as the most relevant predictor, despite a number of exceptions.