title: Characterization and Expression of the Lucina pectinata Oxygen and Sulfide Binding Hemoglobin Genes.
abstract: The clam Lucina pectinata lives in sulfide-rich muds and houses intracellular symbiotic bacteria that need to be supplied with hydrogen sulfide and oxygen. This clam possesses three hemoglobins: hemoglobin I (HbI), a sulfide-reactive protein, and hemoglobin II (HbII) and III (HbIII), which are oxygen-reactive. We characterized the complete gene sequence and promoter regions for the oxygen reactive hemoglobins and the partial structure and promoters of the HbI gene from Lucina pectinata. We show that HbI has two mRNA variants, where the 5'end had either a sequence of 96 bp (long variant) or 37 bp (short variant). The gene structure of the oxygen reactive Hbs is defined by having 4-exons/3-introns with conservation of intron location at B12.2 and G7.0 and the presence of pre-coding introns, while the partial gene structure of HbI has the same intron conservation but appears to have a 5-exon/ 4-intron structure. A search for putative transcription factor binding sites (TFBSs) was done with the promoters for HbII, HbIII, HbI short and HbI long. The HbII, HbIII and HbI long promoters showed similar predicted TFBSs. We also characterized MITE-like elements in the HbI and HbII gene promoters and intronic regions that are similar to sequences found in other mollusk genomes. The gene expression levels of the clam Hbs, from sulfide-rich and sulfide-poor environments showed a significant decrease of expression in the symbiont-containing tissue for those clams in a sulfide-poor environment, suggesting that the sulfide concentration may be involved in the regulation of these proteins. Gene expression evaluation of the two HbI mRNA variants indicated that the longer variant is expressed at higher levels than the shorter variant in both environments.
authors.affiliation: Department of Chemistry, Arts and Sciences Faculty, University of Puerto Rico-Mayaguüez Campus, P.O. Box 9019, Mayagüez, Puerto Rico 00681-9019.
authors.forename: Ingrid M
authors.initials: IM
authors.lastname: Montes-Rodríguez
authors.affiliation: Department of Biochemistry, School of Medicine, University of Puerto Rico-Medical Sciences Campus, P.O. Box 365067, San Juan, Puerto Rico 00936-5067.
authors.forename: Linda E
authors.initials: LE
authors.lastname: Rivera
authors.affiliation: Department of Chemistry, Arts and Sciences Faculty, University of Puerto Rico-Mayaguüez Campus, P.O. Box 9019, Mayagüez, Puerto Rico 00681-9019.
authors.forename: Juan
authors.initials: J
authors.lastname: López-Garriga
authors.affiliation: Department of Biochemistry, School of Medicine, University of Puerto Rico-Medical Sciences Campus, P.O. Box 365067, San Juan, Puerto Rico 00936-5067.
authors.forename: Carmen L
authors.initials: CL
authors.lastname: Cadilla
mesh_terms: D000818:Animals; D001483:Base Sequence; D049872:Bivalvia / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006454:Hemoglobins / Q000235:genetics* / Q000378:metabolism*; D006862:Hydrogen Sulfide / Q000378:metabolism; D008969:Molecular Sequence Data; D010100:Oxygen / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D016415:Sequence Alignment; D013440:Sulfides / Q000378:metabolism*
chemical_list: D006454:Hemoglobins; D012333:RNA, Messenger; D013440:Sulfides; D010100:Oxygen; D006862:Hydrogen Sulfide
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