Identification of a KIR antisense lncRNA expressed by progenitor cells.

Human NK cells express cell surface class I MHC receptors (killer cell immunoglobulin-like receptor, KIR) in a probabilistic manner. Previous studies have shown that a distal promoter acts in conjunction with a proximal bidirectional promoter to control the selective activation of KIR genes. We report here the presence of an intron 2 promoter in several KIR genes that produce a spliced antisense transcript. This long noncoding RNA (lncRNA) transcript contains antisense sequence complementary to KIR-coding exons 1 and 2 as well as the proximal promoter region of the KIR genes. The antisense promoter contains myeloid zinc finger 1 (MZF-1)-binding sites, a transcription factor found in hematopoietic progenitors and myeloid precursors. The KIR antisense lncRNA was detected only in progenitor cells or pluripotent cell lines, suggesting a function that is specific for stem cells. Overexpression of MZF-1 in developing NK cells led to decreased KIR expression, consistent with a role for the KIR antisense lncRNA in silencing KIR gene expression early in development.

The Arabidopsis thaliana ribosomal protein S15 (rig) gene.

We have isolated cDNA and genomic clones for Arabidopsis thaliana cytosolic ribosomal protein S15 and determined their sequences. Like animal S15 genes, this plant S15 gene is composed of four exons and the first intron is located immediately following the ATG translational start codon. The 5' end of the S15 mRNA was mapped by RNase protection experiments which showed that this mRNA contains a 5' untranslated region of approx. 83 nucleotides. Southern blot analyses suggest that Arabidopsis S15 is encoded by a small family of genes. The sequences of the predicted exons in the cloned S15 gene are identical to that of the S15 cDNA, demonstrating that this gene is transcriptionally active. Sequence analysis of the cloned A. thaliana S15 gene shows that it is tightly linked (approx. 500 nucleotides distant) to a gene of unknown function. The Arabidopsis S15 protein described here is about 75% identical to vertebrate S15, 70% identical to the homologous yeast protein (S21), 50% identical to archaebacterial S19, 30% identical to eubacterial S19, and about 30% identical to plant mitochondrial and plastid S19.

Arabidopsis thaliana H1 histones. Analysis of two members of a small gene family.

We have isolated two Arabidopsis thaliana cDNA clones that encodes different H1 histone proteins. The H1-1 and H1-2 proteins are 274 and 273 amino acids in length, respectively. Unlike the H1 histones within a single animal species, the two plant H1 proteins share little sequence similarity outside the protein's central globular domain. Within the globular domain, a pentapeptide that is extremely well conserved in animal H1 histones, is not found in either of the plant proteins. Southern blot analysis suggests that A. thaliana has only three H1 histone genes. A genomic clone encoding the H1-1 protein was isolated and the protein-coding region was found to consist of two exons separated by a 104-bp intron. The site of transcriptional initiation of the H1-1 gene was mapped by primer-extension analysis and a conserved octamer motif, identical to that observed in most plant core histone genes that have been characterized to date, was found 101 nucleotides upstream of the presumed transcription-initiation site. The 3' portion of the gene encoding H1-2 was also isolated and sequenced. When the 3'-flanking regions of the two H1 genes were compared, several highly conserved sequences were observed that might be convergently transcribed relative to the histone genes.

Chlamydomonas transcripts encoding three divergent plastid chaperonins are heat-inducible.

Three cDNAs encoding plastid cpn60 chaperonin subunits have been isolated from the unicellular green alga Chlamydomonas reinhardtii. Based on comparisons of the predicted amino acid sequences, we conclude that Chlamydomonas, like higher plants, contains divergent plastid cpn60-alpha and cpn60-beta subunits. The predicted amino acid sequences of the two Chlamydomonas cpn60-beta subunits differ significantly (24% divergent), indicating that the two cpn60-beta subunits have been selectively maintained for a considerable period of time. Unlike plastid chaperonin transcripts in higher plants, heat shock conditions (42 degrees C) lead to a rapid increase (10- to 30-fold) in the level of each of the three plastid transcripts.

Characterization of rps17, rp19 and rpl15: three nucleus-encoded plastid ribosomal protein genes.

Approximately two-thirds of the 55 to 60 plastid ribosomal proteins are encoded in the nucleus. Since the protein products of each of these genes are needed in equal amounts for ribosome assembly, their expression may be coordinately regulated by common mechanisms. To begin to understand how the expression of these genes is regulated, we have isolated cDNA and genomic clones for three plastid ribosomal protein genes from an Arabidopsis thaliana library. The genes rps17, rpl9 and rpl15, encoding plastid ribosomal proteins CS17, CL9 and CL15, respectively, are located in the nuclear genome and Southern blot data suggest that each is a single copy gene in A. thaliana. Northern blot data show that transcripts from rps17, rpl9 and rpl15 are much more abundant in leaves and stems than they are in roots. The nucleotide sequences of each of these three genes were determined and their transcriptional initiation sites identified. rps17 transcripts have multiple 5' ends suggesting that they are initiated at multiple sites or are post-transcriptionally processed at their 5' end. rpl9 and rpl15 apparently have unique transcriptional initiation sites but are post-transcriptionally processed to remove six and three introns, respectively, from their primary transcripts. We have examined the genomic sequences for motifs that may be important for the proper expression of these genes. A 7 bp sequence motif, whose consensus is 5'-AGGCCCA-3', flanked by AT-rich regions was identified between 38 and 73 nucleotides upstream of the rps17, rpl9 and rpl15 transcriptional initiation sites.