The csmB gene, encoding the 7.5-kDa "Gerola-Olson" protein of chlorosomes, has been cloned and sequenced from the green sulfur bacteria Chlorobium vibrioforme strain 8327D and Chlorobium tepidum. Two potential start codons were identified, and the csmB gene may be translated into a preprotein with an amino-terminal extension. Two forms of the mature CsmB protein (74 or 75 amino acids in length) were identified that differ by the presence or absence of a methionine residue at the amino terminus. The csmB gene of Chl. tepidum is transcribed as an abundant monocistronic mRNA of approximately 350 nucleotides; primer extension mapping of the 5' endpoint of the csmB mRNA suggests there is strong similarity between the csmB promoter and the sigma70 promoters of Escherichia coli. The CsmB protein of Chl. tepidum was overproduced as a histidine-tagged fusion protein in E. coli, purified to homogeneity by Ni2+ chelation affinity chromatography, and used to raise polyclonal antibodies in rabbits. Protease susceptibility mapping and agglutination experiments with isolated chlorosomes using anti-CsmB antibodies indicate that the CsmB protein is a component of the chlorosome envelope.

The csmD and csmE genes, encoding two proteins of the chlorosome envelope, have been cloned and sequenced from the green sulfur bacterium Chlorobium tepidum. The csmD gene predicts a hydrophobic protein of 113 amino acids with a molecular mass of 11.1 kDa. The csmE gene was identified immediately upstream from csmD; the csmE gene predicts a protein of 82 amino acids (9.0 kDa) which is 49% identical to CsmA (Chung et al. (1994) Photosynthesis Res 41: 261-275). The CsmE protein is post-translationally processed, most likely in a manner similar to CsmA. The csmE and csmD genes are cotranscribed as a dicistronic mRNA but can also be cotranscribed with an open reading frame upstream from csmE that predicts a protein with sequence similarity to the CheY and SpoOF subclass of regulatory proteins. The CsmA, CsmC, CsmD, and CsmE proteins were overproduced in Escherichia coli, purified, and used to raise polyclonal antibodies in rabbits. Protease susceptibility mapping and agglutination experiments using these antibodies indicate that all four proteins are exposed at the surface of isolated chlorosomes and hence are probably components of the chlorosome envelope. Additionally, antigalactose antibodies were used to confirm that the galactosyl moiety of monogalactosyl diglycerol is exposed at the chlorosome surface; this is consistent with the notion that these lipids are components of the chlorosome envelope.

The dorsoventral axis of the Drosophila visual cortex is patterned by nonautonomous signals expressed at its dorsal and ventral margins. wingless (wg) expression at the margins induces decapentaplegic (dpp), optomotor blind (omb), and aristaless in adjacent domains. We show that Combgap, a zinc finger protein, represses Wg target gene expression in the visual cortex. Wg signal reception downregulates combgap expression and derepresses target gene transcription. Combgap participates in a Hedgehog-controlled circuit in the developing wing and leg by regulating the expression of Cubitus interruptus. Combgap is thus a tissue-specific relay between Wingless and its target genes for the determination of cell fate in the visual cortex.

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