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In somatic cells the expression of replication-dependent histone genes is coupled to the S phase of the cell cycle. However, we have found a number of novel H2a, H2b, and H3 poly(A)+ RNA species in avian haploid round spermatids. The spermatid-specific H2a and H2b 0.8-kilobase RNAs are transcribed from a subset of the replication-dependent H2a and H2b gene families. Two cDNAs derived from the spermatid-specific H2b transcripts were isolated and sequenced. The structures of these cDNAs reveal that the spermatid-specific RNAs are identical to the 0.5-kilobase poly(A)- H2b mRNAs expressed in proliferating somatic cells, except for the addition of poly(A) at the 3' ends. The site of poly(A) addition in the spermatid-specific RNAs is located 26 to 28 nucleotides 3' of the poly(A)- H2b mRNA terminus. Thus, the hairpin structures and purine-rich elements required for the U7 small nuclear ribonucleoprotein-mediated cleavage reaction that generates the 3' ends of poly(A)- H2b mRNAs are not utilized in spermatids and are retained in the poly(A)+ H2b RNAs.

During mammalian spermiogenesis transitional proteins and protamines replace histones on the DNA as the chromatin condenses. While previous studies suggested that histone genes are inactive postmeiotically, we have shown both by steady-state RNA analysis and nuclear run-off transcription assays that histone 2b (H2b) transcription occurs in mouse round spermatids. In addition, a novel H2b cDNA clone has been isolated from an adult mouse testes cDNA library. The sequence of this cDNA clone predicts a protein that is extremely similar to other mouse H2b proteins, except at the carboxyl-terminus where the testes H2b contains an additional 12 amino acids, seven of which are hydrophobic. In contrast to the replication-dependent histone mRNAs, the 3′ untranslated region of this cDNA contains the poly(A) addition sequence (AAUAAA) upstream of a poly(A) tract. Furthermore, the conserved hairpin structure immediately upstream of replication-dependent histone mRNA termini is not present. Northern blot analysis of RNA from embryonic, ovarian, spermatogenic, and a variety of somatic tissues reveals that this novel H2b transcript is spermatid specific. The H2b mRNA is in polyribosomes isolated from spermatogenic cells, strongly suggesting that it is translated during spermiogenesis.

A simple repetitive DNA sequence in the nuclear genome of Trypanosoma brucei, consisting of tandem repeats of the hexanucleotide 5′ CCCTAA 3′, was identified as being telomeric by several criteria. This sequence was specfically labeled with T. brucei genomic DNA as the template for in vitro nick translation by DNA polymerase I, and was present in Bal 31 nuclease sensitive, genomic restriction fragments of the large sizes expected in this organism for at least some telomeric regions. The same repeated sequence was found in six other flagellates tested. A segment of DNA from T. brucei including this telomeric sequence was cloned in pBR322 and characterized. The cloned segment contained a sequence highly homologous to the 3′ ends of several variant surface glycoprotein mRNAs, upstream of the tandemly repeated hexanucleotide sequence.

The six chicken histone H1 genes have 3'-processing sequences typical of replication-dependent histone genes, which are expressed as poly(A)- mRNAs. However, by Northern analysis of RNA from several adult chicken tissues, as well as from embryonal skeletal muscle in vivo and in vitro, we have observed histone H1 transcripts longer than those predicted on the basis of the published genomic sequences. These RNAs are polyadenylated transcripts of the genes H1.01 and H1.10, which encode the 'c fraction' H1 protein subtypes. Both transcripts contain an internal stem-loop and purine-rich box associated with the 3' processing of poly(A)- histone mRNAs. The 2-kb poly(A)+ H1.01 transcript is present at high steady-state levels in tissues with low rates of DNA synthesis, has a longer half-life than the poly(A)- mRNA from the same gene, and is polyribosomal in embryonal skeletal muscle. The 1-kb poly(A)+ H1.10 RNA is the major H1.10 transcript in adult skeletal muscle. The properties of these RNAs suggest that they may contribute to the relaxed replication dependence of c fraction subtype expression. The polyadenylation signals of both genes are unusual in their association with processed (nonhistone) pseudogene-like elements, an arrangement with possible implications for the mechanism of alternative 3'-end formation in these genes.