Bioinformatic analysis of Ci proteins of Apis mellifera

【Objective】To apprehend bioinformatics of transcription factor Apis mellifera(Ci) and elucidate its functions,and to lay a foundation for exploring functions and roles of Ci proteins in Hh signaling pathway of A. mellifera.【Method】Ci protein amino acid sequences of A. mellifera were obtained from NCBI,and their physicochemical properties were predicted using ProtParam,predicting signal peptides by SignalP-5.0,transmembrane structures by TMHMM-2.0,O-glycosylation sites by NetOGlyc 3.0,N-glycosylation sites by NetNGlyc 1.0,phosphorylation sites by NetPhos 3.1,sumoylation sites by SUMOplot and tertiary structure of Ci proteins in A. mellifera by GOR4,SWISS-MODEL,CDSearch. Based on multiple sequence alignment,phylogenetic trees were constructed by MEGA 11.0 and interacting proteins were predicted by String database.【Result】Ci proteins in A. mellifera had two subtypes,XP_624136.4 and XP_006558245.2. XP_624136.4 contained open reading frames(ORF) of 4338 bp,encoding 1445 amino acid residues,encoding protein molecular mass of 15.50 kD and a theoretical isoelectric point(pI) of 8.39;XP_006558245.2 contained ORF of 3873 bp,encoding 1290 amino acid residues,encoding protein molecular mass of 13.99 kD and a pI of 8.48. Both were unstable amphiphilic proteins with no signal peptide or transmembrane structure,and they localized mainly in nucleus and a few of them in the vesicle cytoplasm. A few of XP_006558245.2 localized in mitochondria. XP_624136.4 had 2O-glycosylation sites,9 N-glycosylation sites,174 phosphorylation sites and 5 hematoxylation sites;XP_006558245.2had 26 O-glycosylation sites,8 N-glycosylation sites,156 phosphorylation sites and 5 hematoxylation sites. In secondary structures of Ci proteins A. mellifera were mainly α-helix,folded extended chains and irregular curls;in tertiary structures of the protein were mainly irregular curls and extended chains and a few α-helix;both subtypes had 5 typical C₂H₂-type zinc finger protein structural domains with highly conserved Ci protein sequences from insects to mammals. Ci proteins in A. mellifera and kinesin-like proteins such as Kinesin-B,Ptc,Poz,Su(fu),Slmb,Smo,Csnk1a1,Cul-3 and Fu proteins formed an interaction network.【Conclusion】Ci proteins in A. mellifera are unstable amphiphilic proteins,mainly localize in the nucleus,and a few distribute in vesicle cytoplasm or mitochondria. The proteins have 5 typical C₂H₂-type zinc finger protein structural domains and highly conserved protein sequences,which mainly work for transcription in Hh signaling pathway of A. mellifera,probably play an important role in regulation of A. mellifera growth and development,transmembrane transport,synaptic transmission,signal transduction and protein production.