Differential expression of Homeobox C11 protein in water buffalo Bubalus bubalis and its putative 3D structure

被引:0
作者
Sharma, Monal [1 ]
Rawal, Leena [1 ]
Panwar, Deepak [1 ]
Sehgal, Neeta [2 ]
Ali, Sher [1 ]
机构
[1] Natl Inst Immunol, Mol Genet Lab, New Delhi 110067, India
[2] Univ Delhi, Dept Zool, Delhi 110007, India
来源
BMC GENOMICS | 2014年 / 15卷
关键词
HOXC11; protein; Water buffalo; Gene expression; Protein characterization; Protein modeling; HOX GENES; LACTASE; HOMEODOMAIN;
D O I
10.1186/1471-2164-15-638
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Background: The Homeobox (Hox) family complex contains 39 genes, clustered into four groups (A-D) all expressing in sequential manner. The HOX proteins are transcriptional factors involved in regulation of pattern formation of the anterio-posterior body axis across the species. Most of the Hox family genes have been studied with respect to their organization and expression during the embryonic stages. However, expression pattern of Homeobox C11 (Hoxc11) gene in the 5' region, particularly in higher mammals remains largely unexplored. Results: We cloned and expressed Homeobox C11 (Hoxc11) gene from water buffalo Bubalus bubalis. The recombinant HOXC11 protein expressed as inclusion bodies was solubilized in Tris buffer (10 mM, pH-6.5) and purified using Ni-NTA affinity column. The purity and molecular weight of HOXC11 protein (similar to 33 kDa) were confirmed by SDS-PAGE and western blot analysis. Employing immunohistochemistry approach, we localized HOXC11 protein in the nuclei across the tissues of buffalo. Western blot analysis showed highest expression of HOXC11 protein in kidney and lung although its possible renal and respiratory roles are not yet established. Electrophoretic mobility shift assay (EMSA) demonstrated the specific binding of HOXC11 protein with the promoter element, CE-LPH1 of lactase-phlorizin hydrolase (LPH) gene showing reduced mobility of the protein DNA complex, corroborating with earlier report on the possible role of this protein in intestinal functions. In silico analysis of HOXC11 showed predominance of a helices and presence of six conserved domains. We deduced the putative 3D structure of HOXC11 protein and fifteen possible DNA interacting residues within the homeodomain. Conclusions: Present study augments our understanding on the specific expression of HOXC11 protein in kidney and lung in water buffalo. The fifteen DNA interacting residues reported herein provide an opportunity to establish much broader structural and functional perspectives of HOXC11 protein in the context of genome analysis in general and animal biotechnology in particular.
引用
收藏
页数:11
相关论文
共 42 条
  • [1] Transcription of homeobox-containing genes detected in cDNA libraries derived from human unfertilized oocytes and preimplantation embryos
    Adjaye, J
    Monk, M
    [J]. MOLECULAR HUMAN REPRODUCTION, 2000, 6 (08) : 707 - 711
  • [2] [Anonymous], SCAND J NUTR
  • [3] Increased HOX C13 expression in metastatic melanoma progression
    Cantile, Monica
    Scognamiglio, Giosue
    Anniciello, Annamaria
    Farina, Marisa
    Gentilcore, Giusy
    Santonastaso, Clemente
    Fulciniti, Franco
    Cillo, Clemente
    Franco, Renato
    Ascierto, Paolo A.
    Botti, Gerardo
    [J]. JOURNAL OF TRANSLATIONAL MEDICINE, 2012, 10
  • [4] VERIFICATION OF PROTEIN STRUCTURES - PATTERNS OF NONBONDED ATOMIC INTERACTIONS
    COLOVOS, C
    YEATES, TO
    [J]. PROTEIN SCIENCE, 1993, 2 (09) : 1511 - 1519
  • [5] Developmental functions of mammalian Hox genes
    Favier, B
    Dolle, P
    [J]. MOLECULAR HUMAN REPRODUCTION, 1997, 3 (02) : 115 - 131
  • [6] FRIEDMANN Y, 1994, CANCER RES, V54, P5981
  • [7] Regulation of H3K27me3 and H3K4me3 During Early Porcine Embryonic Development
    Gao, Yu
    Hyttel, Poul
    Hall, Vanessa Jane
    [J]. MOLECULAR REPRODUCTION AND DEVELOPMENT, 2010, 77 (06) : 540 - 549
  • [8] GEHRING WJ, 1994, ANNU REV BIOCHEM, V63, P487, DOI 10.1146/annurev.bi.63.070194.002415
  • [9] Limb malformations and the human HOX genes
    Goodman, FR
    [J]. AMERICAN JOURNAL OF MEDICAL GENETICS, 2002, 112 (03): : 256 - 265
  • [10] Using circular dichroism spectra to estimate protein secondary structure
    Greenfield, Norma J.
    [J]. NATURE PROTOCOLS, 2006, 1 (06) : 2876 - 2890