Bioinformatic Identification of Plant Hydroxyproline-Rich Glycoproteins.

Hydroxyproline-rich glycoproteins (HRGPs) are a superfamily of plant cell wall proteins that function in diverse aspects of plant growth and development. This superfamily consists of three members: arabinogalactan-proteins (AGPs), extensins (EXTs), and proline-rich proteins (PRPs). Hybrid and chimeric HRGPs also exist. A bioinformatic software program, BIO OHIO 2.0, was developed to expedite the genome-wide identification and classification of AGPs, EXTs, and PRPs based on characteristic HRGP motifs and biased amino acid compositions. This chapter explains the principles of identifying HRGPs and provides a stepwise tutorial for using the BIO OHIO 2.0 program with genomic/proteomic data. Here, as an example, the genome/proteome of the common bean (Phaseolus vulgaris) is analyzed using the BIO OHIO 2.0 program to identify and characterize its set of HRGPs.

Expression of arabinogalactan proteins during tomato fruit ripening and in response to mechanical wounding, hypoxia and anoxia.

Arabinogalactan proteins (AGPs) are highly glycosylated members of the superfamily of hydroxyproline-rich glycoproteins (HRGPs). Despite their implication in many aspects of plant growth and development little is known about their role in tomato fruit ripening (Solanum lycopersicum) and their response to abiotic stress in tomato fruits. A search of the currently available tomato genome database resulted in the identification of 34 genes encoding putative AGPs, with at least 20 of them being expressed in fruit. We monitored the abundance of AGPs bound by JIM8 and JIM13 monoclonal antibodies as well as the gene expression profiles of the Lys-rich LeAGP1 and two classical AGPs, SlAGP2 and SlAGP4. The JIM8- and JIM13-bound AGPs showed constitutive expression during fruit ripening and under hypoxic conditions, slight up-regulation to mechanical wounding in excised tomato fruit pericarp discs and up-regulation under anoxia indicating functional roles for these proteins in the developmental program of ripening and in response to abiotic stresses. Moreover, the SlAGP2 mRNA was significantly up-regulated during fruit ripening following the climacteric ethylene production, a pattern of expression similar to that of tomato fruit PG. The SlAGP4 and LeAGP1 mRNAs were up-regulated in response to mechanical wounding while under anoxia only the SlAGP4 transcript was induced. The protein and mRNA levels of these AGPs were induced under mechanical wounding while only JIM8-bound AGPs and SIAGP4 expression were induced under anoxic conditions. Our results indicate that selected tomato AGPs seem to play a role in fruit ripening as well as in response to mechanical wounding and anoxia.

Genome-wide identification, classification, and expression analysis of the arabinogalactan protein gene family in rice (Oryza sativa L.).

Arabinogalactan proteins (AGPs) comprise a family of hydroxyproline-rich glycoproteins that are implicated in plant growth and development. In this study, 69 AGPs are identified from the rice genome, including 13 classical AGPs, 15 arabinogalactan (AG) peptides, three non-classical AGPs, three early nodulin-like AGPs (eNod-like AGPs), eight non-specific lipid transfer protein-like AGPs (nsLTP-like AGPs), and 27 fasciclin-like AGPs (FLAs). The results from expressed sequence tags, microarrays, and massively parallel signature sequencing tags are used to analyse the expression of AGP-encoding genes, which is confirmed by real-time PCR. The results reveal that several rice AGP-encoding genes are predominantly expressed in anthers and display differential expression patterns in response to abscisic acid, gibberellic acid, and abiotic stresses. Based on the results obtained from this analysis, an attempt has been made to link the protein structures and expression patterns of rice AGP-encoding genes to their functions. Taken together, the genome-wide identification and expression analysis of the rice AGP gene family might facilitate further functional studies of rice AGPs.

Identification of a novel group of putative Arabidopsis thaliana beta-(1,3)-galactosyltransferases.

To begin biochemical and molecular studies on the biosynthesis of the type II arabinogalactan chains on arabinogalactan-proteins (AGPs), we adopted a bioinformatic approach to identify and systematically characterise the putative galactosyltransferases (GalTs) responsible for synthesizing the beta-(1,3)-Gal linkage from CAZy GT-family-31 from Arabidopsis thaliana. These analyses confirmed that 20 members of the GT-31 family contained domains/motifs typical of biochemically characterised beta-(1,3)-GTs from mammalian systems. Microarray data confirm that members of this family are expressed throughout all tissues making them likely candidates for the assembly of the ubiquitously found AGPs. One member, At1g77810, was selected for further analysis including location studies that confirmed its presence in the Golgi and preliminary enzyme substrate specificity studies that demonstrated beta-(1,3)-GalT activity. This bioinformatic/molecular study of CAZy GT-family-31 was validated by the recent report of Strasser et al. (Plant Cell 19:2278-2292, 2007) that another member of this family (At1g26810; GALT1) encodes a beta-(1,3)-GalT involved in the biosynthesis of the Lewis a epitope of N-glycans in Arabidopsis thaliana.

Characterization of 19 novel cotton FLA genes and their expression profiling in fiber development and in response to phytohormones and salt stress.

Fasciclin-like arabinogalactan proteins (FLAs), a subclass of arabinogalactan proteins (AGPs), are usually involved in cell development in plants. To investigate the expression profiling as well as the role of FLA genes in fiber development, 19 GhFLA genes (cDNAs) were isolated from cotton (Gossypium hirsutum). Among them, 15 are predicted to be glycosylphosphatidylinositol anchored to the plasma membranes. The isolated cotton FLAs could be divided into four groups. Real-time quantitative reverse transcriptase polymerase chain reaction results indicated that the GhFLA genes are differentially expressed in cotton tissues. Three genes (GhFLA1/2/4) were specifically or predominantly expressed in 10 days post-anthesis fibers, and the transcripts of the other four genes (GhFLA6/14/15/18) were accumulated at relatively high levels in cotton fibers. Furthermore, expressions of the GhFLA genes are regulated in fiber development and in response to phytohormones and NaCl. The identification of cotton FLAs will facilitate the study of their roles in cotton fiber development and cell wall biogenesis.

Duplication and selection on abalone sperm lysin in an allopatric population.

While gene duplication is a major source of evolutionary novelty, the importance of this process in reproductive protein evolution has not been widely investigated. Here, we report the first known case of gene duplication of abalone sperm lysin in an allopatric subspecies found in the Eastern Atlantic, Haliotis tuberculata coccinea. Mass spectrometry identified both copies of the lysin protein in testis tissue, and 3-dimensional structural modeling suggests that both proteins remain functional. We also detected positive selection acting on both paralogs after duplication and found evidence of a recent selective sweep. Because H. t. coccinea occurs in geographic isolation from other abalone species, these findings suggest that the evolution of lysin is not driven to create reproductive barriers to unfit hybrid formation with an overlapping species. Instead, sexual selection or sexual conflict acting during abalone fertilization could be responsible for the recent positive selection on this protein. The presence of multiple, rapidly evolving lysin genes in H. tuberculata presents an opportunity to study the early stages of diversification of a protein whose function is well understood.

Niveles séricos de mucoproteínas en escolares sanos residentes en la ciudad de Mérida: Venezuela / Mucoproteins serum levels in schools children from Mérida: Venezuela

El interés sobre las glicoproteínas séricas, incluyendo las mucoproteínas, se reflejan en la gran cantidad de publicaciones que existen sobre el tema. La mayoría de ellas se refiere a personas adultas no encontrándose ningún estudio completo sobre estos compuestos durante la niñez y la adolescencia. En la presente investigación se cuantificó el contenido sérico de las mucoproteínas en 320 escolares en buen estado de salud:160 niños y 160 niñas, entre 7 y 14 años de edad, procedentes de la ciudad de Mérida(Venezuela). Las mucoproteínas alcanzaron un promedio de 15ñ4mg/dl(rango=88-18), no demostrándose ninguna variación significativa en este valor en relación con la edad o con el sexo de los escolares sometidos al estudio