title: A prawn core histone 4: derivation of N- and C-terminal peptides and their antimicrobial properties, molecular characterization and mRNA transcription.
abstract: This study investigates the complete molecular characterization including bioinformatics characterization, gene expression, synthesis of N and C terminal peptides and their antimicrobial activity of the core histone 4 (H4) from freshwater giant prawn Macrobrachium rosenbergii (Mr). A cDNA encoding MrH4 was identified from the constructed cDNA library of M. rosenbergii during screening and the sequence was obtained using internal sequencing primers. The MrH4 coding region possesses a polypeptide of 103 amino acids with a calculated molecular weight of 11kDa and an isoelectric point of 11.5. The bioinformatics analysis showed that the MrH4 polypeptide contains a H4 signature at (15)GAKRH(19). Multiple sequence alignment of MrH4 showed that the N-terminal (21-42) and C-terminal (87-101) antimicrobial peptide regions and the pentapeptide or H4 signature (15-19) are highly conserved including in humans. The phylogenetic tree formed two separate clades of vertebrate and invertebrate H4, wherein MrH4 was located within the arthropod monophyletic clade of invertebrate H4 groups. Three-dimensional model of MrH4 was established using I-TASSER program and the model was validated using Ramachandran plot analysis. Schiffer-Edmundson helical wheel modeling was used to predict the helix propensity of N (21-42) and C (87-101) terminal derived Mr peptides. The highest gene expression was observed in gills and is induced by viral [white spot syndrome baculovirus (WSBV) and M. rosenbergii nodovirus (MrNV)] and bacterial (Aeromonas hydrophila and Vibrio harveyi) infections. The N and C terminal peptides were synthesized and their antimicrobial and hemolytic properties were examined. Both peptides showed activity against the tested Gram negative and Gram positive bacteria; however, the highest activity was noticed against Gram negative bacteria. Among the two peptides used in this study, C-terminal peptide yielded better results than the N-terminal peptide. Therefore, C terminal peptide can be recommended for the development of an antimicrobial agent.
authors.affiliation: Division of Fisheries Biotechnology and Molecular Biology, Research Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, Chennai 603203, Tamil Nadu, India.
authors.forename: Mukesh Kumar
authors.initials: MK
authors.lastname: Chaurasia
authors.affiliation: Division of Fisheries Biotechnology and Molecular Biology, Research Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, Chennai 603203, Tamil Nadu, India.
authors.forename: Rajesh
authors.initials: R
authors.lastname: Palanisamy
authors.affiliation: Division of Fisheries Biotechnology and Molecular Biology, Research Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, Chennai 603203, Tamil Nadu, India.
authors.forename: Prasanth
authors.initials: P
authors.lastname: Bhatt
authors.affiliation: Division of Fisheries Biotechnology and Molecular Biology, Research Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, Chennai 603203, Tamil Nadu, India.
authors.forename: Venkatesh
authors.initials: V
authors.lastname: Kumaresan
authors.affiliation: Institute for Cellular and Molecular Biology, The University of Texas at Austin, 1 University Station A4800, Austin, TX 78712, USA.
authors.forename: Annie J
authors.initials: AJ
authors.lastname: Gnanam
authors.affiliation: Lab PCN 206, Microbiology Division, CSIR-Central Drug Research Institute, B.S. 10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow 226031, Uttar Pradesh, India.
authors.forename: Mukesh
authors.initials: M
authors.lastname: Pasupuleti
authors.affiliation: Department of Biotechnology, Faculty of Applied Sciences, AIMST University, Semeling Bedong, 08100 Bedong, Kedah, Malaysia.
authors.forename: Marimuthu
authors.initials: M
authors.lastname: Kasi
authors.affiliation: PG and Research Department of Biotechnology, Bharath College of Science and Management, Thanjavur 613 005, Tamil Nadu, India.
authors.forename: Ramaswamy
authors.initials: R
authors.lastname: Harikrishnan
authors.affiliation: Division of Fisheries Biotechnology and Molecular Biology, Research Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, Chennai 603203, Tamil Nadu, India. Electronic address: jesuaraj@hotmail.com.
authors.forename: Jesu
authors.initials: J
authors.lastname: Arockiaraj
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000890:Anti-Infective Agents / Q000737:chemistry / Q000494:pharmacology; D001483:Base Sequence; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D015870:Gene Expression; D006461:Hemolysis / Q000187:drug effects; D006657:Histones / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D033541:Palaemonidae; D010455:Peptides / Q000737:chemistry / Q000494:pharmacology; D010802:Phylogeny; D017433:Protein Structure, Secondary; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment; D014158:Transcription, Genetic
chemical_list: D000890:Anti-Infective Agents; D018076:DNA, Complementary; D006657:Histones; D010455:Peptides; D012333:RNA, Messenger
keywords: Antimicrobial peptide; Gene expression; Hemolysis; Histone; Prawn