Jinhu Guo

Cloning and identification of the human LPAAT-zeta gene, a novel member of the lysophosphatidic acid acyltransferase family

AbstractLysophosphatidic acid (LPA) is a naturally occurring component of phospholipid and plays a critical role in the regulation of many physiological and pathophysiological processes including cell growth, survival, and pro-angiogenesis. LPA is converted to phosphatidic acid by the action of lysophosphatidic acid acyltransferase (LPAAT). Five members of the LPAAT gene family have been detected in humans to date. Here, we report the identification of a novel LPAAT member, which is designated as LPAAT-ζ. LPAAT-ζ was predicted to encode a protein consisting of 456 amino acid residues with a signal peptide sequence and the acyltransferase domain. Northern blot analysis showed that LPAAT-ζ was ubiquitously expressed in all 16 human tissues examined, with levels in the skeletal muscle, heart, and testis being relatively high and in the lung being relatively low. The human LPAAT-ζ gene consisted of 13 exons and is positioned at chromosome 8p11.21.

In silico analysis indicates a similar gene expression pattern between human brain and testis

Objectives: Previous data has reported similarity between human brain and testis gene expression patterns. Brain is the most important tissue in human speciation. Hence, it means that human testis could also play a crucial role in human speciation if these two tissues exhibit similar gene expression patterns. However, previous reports were based on only limited and scattered data. Determining the large scale anatomy of gene expression patterns of various human tissues could draw a more convincing conclusion, and better our understanding of the correlation/inter-correlation among different tissues. Furthermore, it could also provide a clue for evolutionary study. Methods: To obtain gene expression information for large-scale data analysis, expression data of 760 Unigenes in seventeen human tissues (liver, lung, testis, brain, ovary, uterus, colon, stomach, heart, eye, kidney, spleen, gall bladder, breast, thymus, prostate and pancreas) were retrieved by DDD (differential digital display) analysis, and this expression data was subjected to clustering analysis. These Unigenes represent a wide range of genes classified according to their characterization and function. Results: Among the 17 tissues, the highest similarity in gene expression patterns was between human brain and testis, based on DDD and clustering analysis. Genes contributing to the similarity include ribosomal protein (RP) genes as well as genes involved in transcription, translation and cell division. Conclusions: Present results provide evidence to support the proposal that human testis and brain share the highest similarity of gene expression patterns. The implications of the similarity regarding that both brain and testis contributed to human speciation are discussed.

Cloning and characterization a novel human 1-acyl-sn-glycerol-3-phosphate acyltransferase gene AGPAT7.

The 1-Acylglycerolphosphate acyltransferase is crucial enzyme for synthesis of glycerolipids as well as triacylglylcerol biosynthesis in eukaryotes. Six members of 1-acyl-sn-glycerol-3-phosphate acyltransferase family in human have been described, which were AGPAT1, 2, 3, 4, 5 and 6. Here we report the cloning and characterization of another novel human 1-acyl-sn-glycerol-3-phosphate acyltransferase member AGPAT7 (1-acyl-sn-glycerol-3-phosphate acyltransferase 7) gene, which was mapped to human chromosome 15q14. The AGPAT7 cDNA is 1898 bp in length, encoding a putative protein with 524 amino acid residues, which contains an acyltransferase domain in 123–234 aa. RT PCR amplification in 18 human tissues indicated that human AGPAT7 gene was widely expressed in uterus, thymus, pancreas, skeletal muscle, bladder, stomach, lung and testis. AGPAT7 protein was mainly localized to the endoplasmic reticulum (ER) in Hela cells.

Transcriptomic analyses support the similarity of gene expression between brain and testis in human as well as mouse

We previously revealed similarity in gene expression patterns between human brain and testis, based on digital differential display analyses of 760 human Unigenes. In the present work, we reanalyzed the gene expression data in many tissues of human and mouse for a large number of genes almost covering the respective whole genomes. The results indicated that both in human and in mouse, the gene expression profiles exhibited by brain, cerebellum and testis are most similar to each other compared with other tissues.

Identification and characterization of D8C, a novel domain present in liver‐specific LZP, uromodulin and glycoprotein 2, mutated in familial juvenile hyperuricaemic nephropathy

Present work reported a novel domain – D8C (domain with conserved eight cysteines in liver‐specific ZP domain‐containing protein, glycoprotein 2 (GP‐2) and uromodulin (UMOD)), present in liver‐specific LZP, UMOD, GP‐2 and some uncharacterized proteins, most of which are membrane proteins, extracellular proteins or nuclear membrane proteins. D8C contains eight well‐conserved cysteine residues, which were predicted to form four pairs of disulfide bridges. D8C is composed mainly of β‐strands. Mutation in the D8C at Cys217 in human UMOD is associated with familial juvenile hyperuricaemic nephropathy, which might be due to the disruption of the disulfide bridge. Identification of D8C would further the understandings of related proteins.

Natural antisense transcripts of Alzheimer's disease associated genes.

Natural antisense transcripts (NATs), also named endogenous antisense transcripts, are a class of genes whose role in controlling gene expression is becoming more and more relevant. NATs might play important roles in gene expression and translation regulation. Present work investigated the presence of NATs of Alzheimers disease associated genes including PRESENILIN1, PRESENILIN2, BACE1, BACE2, APP, APOE, TAU (MAPT), PRION, α-SYNUCLEIN (SNCA), NICASTRIN, PEN2, APH1A, APH1B as well as CD147 (BASIGIN), and the results revealed that APP, BACE2, APH1A, TAU, CD147 and α-SYNUCLEIN contain natural antisense transcripts. These NATs were characterized according to the sense–antisense overlapping information and potential functional mechanisms were proposed. Present findings provide preliminary but important information about transcription regulation of AD associated genes, which would further our understanding of the gene expression regulation of AD, and also suggest a novel potential strategy for the therapy of AD.

Isolation and characterization of the human d-glyceric acidemia related glycerate kinase gene GLYCTK1 and its alternatively splicing variant GLYCTK2

Deficiency of human glycerate kinase leads to d-glycerate acidemia/d-glyceric aciduria. Through PCR cloning assisted by in silico approach, we isolated the human glycerate kinase genes—Glycerate Kinase 1 (GLYCTK1) and its alternatively splicing variant—Glycerate Kinase 2 (GLYCTK2), which might be associated with d-glycerate acidemia/d-glyceric aciduria. The locus of GLYCTK gene is mapped to 3p21. PCR amplification in seventeen human tissue cDNAs revealed that both GLYCTK1 and GLYCTK2 are expressed widely almost in all these tissues. The expression of mouse Glyctk in various tissues was demonstrated by in situ hybridization. Both GLYCTK1 and GLYCTK2 proteins are localized in cytosol, and GLYCTK2 proteins are specifically localized in mitochondria. Present results revealed the characteristic expression pattern of murine Glyctk in neural system, skeleton muscle, supporting that glycerate kinase is implicated in d-glycerate acidemia/d-glyceric aciduria.

Cloning and Characterization of Human CAGLP Gene Encoding a Novel EF-hand Protein

The EF-hand proteins, containing conserved Ca2+ binding motifs, play important roles in many biological processes. Through data mining, a novel human gene, CAGLP (calglandulin-like protein) was predicted and subsequently isolated from human skeleton muscle. The open reading frame of CAGLP is 543 bp in length, coding a putative Ca2+ binding protein with four EF-hand motifs. The deduced amino acid sequence of CAGLP displays high similarity with Bothrops insularis snake protein calglandulin (80%). The results of PCR amplification using cDNA from 17 human tissues indicated that human CAGLP is expressed in prostate, thymus, heart, skeleton muscle, bone marrow and ovary. Functional CAGLP::EGFP (enhanced green fluorescent protein) fusion protein revealed that CAGLP accumulated through-out Hela cells. Western blot using anti-EGFP antibodies indicated that the CAGLP protein has a molecular weight of about 19 kD. A phylogenetic tree showed that CAGLP and calglandulin may be orthologous proteins representing a distinct group in the EF-hand proteins.

EFCBP1/NECAB1, a brain-specifically expressed gene with highest abundance in temporal lobe, encodes a protein containing EF-hand and antibiotic biosynthesis monooxygenase domains

Human EFCBP/NECAB family consists of important participants in neuronal calcium signaling, including EFCBP1/NECAB1, EFCBP2/NECAB2 and EFCBP3/NECAB3. In the present study, we identified the full-length 5229 bp EFCBP1 cDNA which was not described before. Human EFCBP1 encodes a 351 amino acid protein containing two EF-hand motifs and an antibiotic biosynthesis monooxygenase (ABM) domain, sharing 49.9 and 56.8% global homology with human EFCBP2 and EFCBP3. Northern hybridization revealed that EFCBP1 is specifically expressed in brain and its abundance varies in different brain regions. EFCBP1s abundance in temporal lobe, frontal lobe and occipital pole is about 3.4, 1.9 and 1.5 folds of the average abundance in cerebral cortex, respectively. The expression level of EFCBP1 equals in putamen and cerebral cortex, while no hybridization signal was detected in spinal cord. In addition, we found that EFCBP1, EFCBP2 and EFCBP3 share a similar exon distribution mode, though their chromosomal localizations, genomic sizes and intron sizes are diverse.

Expression pattern of growth/differentiation factor 3 in human and murine cerebral cortex, hippocampus as well as cerebellum

Growth/differentiation factor 3 is a member of GDF/BMP subfamily of the TGF-beta superfamily, which has been reported to be implicated in testis carcinoma and deposition of adipose tissue. Interestingly, present work indicated that GDF3/Gdf3 genes were expressed in cerebral cortex, hippocampus as well as in cerebellum, as revealed by RT-PCR, in situ hybridization and immunostaining. Results of RT-PCR in 10 human tissues and 12 rat tissues indicated that GDF3/Gdf3 genes were abundantly transcribed in both human and murine brain, including cerebral cortex, hippocampus and cerebellum. In situ hybridization and immunohistochemistry results revealed that in cerebral cortex, GDF3 was evenly distributed. In hippocampus, it was expressed in most of the neurons in CA2 and DG region, especially only in a restricted number of neurons in the regions of CA1 and CA3 and in Purkinje cells in cerebellum. Present data suggested that GDF3 might play important roles in the central nervous system (CNS), especially in cerebral cortex, hippocampus and cerebellum, and it shed new light on further research of GDF3 in the central nervous system.