Two proteins crosslinked to RNA containing the adenovirus L3 poly(A) site require the AAUAAA sequence for binding.

Moore, Claire; Chen, Jie; Whoriskey, John

doi:10.1002/j.1460-2075.1988.tb03183.x

Cited by 114 publications

(100 citation statements)

References 61 publications

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“…Deletion of nucleotides beyond the cleavage site reduces the ability of CPSF-160 to bind to the PAS. Mutations of the PAS abolished the cross-linking of nuclear extract [89], while they only have a small effect on the cross-linking of the recombinant CPSF-160 [88], suggesting that other proteins are required for the highly specific binding to the PAS by CPSF-160. CPSF-160 can also bind at a site other than the PAS in HIV pre-mRNA [90].…”

Section: Cpsf-160 (Cft1p/yhh1p)mentioning

confidence: 99%

Protein factors in pre-mRNA 3′-end processing

Mandel

Bai

Tong

2007

Cell. Mol. Life Sci.

364

482

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Most eukaryotic mRNA precursors (pre-mRNAs) must undergo extensive processing, including cleavage and polyadenylation at the 3′-end. Processing at the 3′-end is controlled by sequence elements in the pre-mRNA (cis elements) as well as protein factors. Despite the seeming biochemical simplicity of the processing reactions, more than 14 proteins have been identified for the mammalian complex, and more than 20 proteins have been identified for the yeast complex. The 3′-end processing machinery also has important roles in transcription and splicing. The mammalian machinery contains several sub-complexes, including cleavage and polyadenylation specificity factor (CPSF), cleavage stimulation factor (CstF), cleavage factor I (CF I m ), and cleavage factor II (CF II m ). Additional protein factors include poly(A) polymerase (PAP), poly(A) binding protein (PABP), symplekin, and the C-terminal domain (CTD) of RNA polymerase II largest subunit. The yeast machinery includes cleavage factor IA (CF IA), cleavage factor IB (CF IB), and cleavage and polyadenylation factor (CPF).

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Section: Cpsf-160 (Cft1p/yhh1p)mentioning

confidence: 99%

Protein factors in pre-mRNA 3′-end processing

Mandel

Bai

Tong

2007

Cell. Mol. Life Sci.

364

482

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“…Alternatively, it may be the subunit in contact with the cleavage site, or it may interact with both sites. The first possibility would be reminiscent of early studies which showed that the binding of the p64 subunit of mammalian CstF was dependent on the AAUAAA sequence (37,38).…”

Section: Table Imentioning

confidence: 99%

Cleavage Factor II of Saccharomyces cerevisiaeContains Homologues to Subunits of the Mammalian Cleavage/ Polyadenylation Specificity Factor and Exhibits Sequence-specific, ATP-dependent Interaction with Precursor RNA

Zhao

Kessler

Moore

1997

Journal of Biological Chemistry

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Cleavage of pre-mRNA during 3 -end formation in yeast requires two protein factors, cleavage factor I (CF I) and cleavage factor (CF II). A 5300-fold purification of CF II indicates that four polypeptides of 150, 105, 100, and 90 kDa copurify with CF II activity. The 150-kDa protein is recognized by antibodies against Cft1, the yeast homologue of the 160-kDa subunit of the mammalian cleavage/polyadenylation specificity factor (CPSF). The 100-kDa subunit is identical to Brr5/Ysh1, a yeast protein with striking similarity to the 73-kDa subunit of CPSF. The 105-kDa protein, designated Cft2 (cleavage factor two) exhibits significant homology to the CPSF 100-kDa subunit. Cft2 is cross-linked to pre-mRNA substrate containing the poly(A) site and wild type upstream and downstream flanking sequences, but not to precleaved RNA lacking downstream sequences or to substrate in which the (UA) 6 processing signal has been deleted. The specific binding of Cft2 to the RNA substrate is ATP-dependent, in agreement with the requirement of ATP for cleavage. The sequence-specific binding of Cft2 and the similarities of CF II subunits to those of CPSF supports the hypothesis that CF II functions in the cleavage of yeast mRNA 3 -ends in a manner analagous to that of CPSF in the mammalian system. These results provide additional evidence that certain features of the molecular mechanism of mRNA 3 -end formation are conserved between yeast and mammals, but also highlight unexpected differences.The formation of messenger RNA 3Ј-ends, an important step in maturation of eukaryotic mRNAs, requires two events, sitespecific endonucleolytic cleavage of primary transcripts followed by poly(A) addition to the upstream fragment. While cleavage and polyadenylation are closely coupled in vivo, the two reactions can be experimentally uncoupled and assayed separately, allowing biochemical characterization of individual components. The 3Ј-end processing of mRNA depends on both cis-acting elements and trans-acting protein factors (for reviews, see Refs. 1-4). Six protein factors act in concert to recognize, cleave, and polyadenylate mammalian pre-mRNAs. (6, 7), and the CstF binds to the GUor U-rich downstream element via its 64-kDa subunit (8).In the yeast Saccharomyces cerevisiae, three cis-acting elements are thought to be necessary and sufficient for mRNA 3Ј-end formation (Refs. 9 and 10 and references therein): the efficiency element formed by a UA repeat or related sequences, which functions by enhancing the efficiency of the positioning element; the positioning element, formed by AAUAAA or related sequences, which positions the cleavage site; and the actual cleavage site, a Py(A) n sequence. Both the efficiency and the positioning elements are found upstream of the poly(A) site.Four functionally distinct factors are required for specific cleavage and polyadenylation of yeast pre-mRNA in vitro (11). Cleavage factors I and II (CF I and CF II) are sufficient for the cleavage reaction, while specific poly(A) addition needs CF I, PAP, and polyadenylat...

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“…While examining these differences, we discovered CstF-64 (8), which is a paralog of the 64,000 M r subunit of the cleavage stimulation factor (CstF-64) (9)(10)(11). CstF-64 is expressed in nuclei of early spermatogenic cells (5,12).…”

mentioning

confidence: 99%

Loss of polyadenylation protein τCstF-64 causes spermatogenic defects and male infertility

Dass

Tardif

Park

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

131

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Polyadenylation, the process of eukaryotic mRNA 3 end formation, is essential for gene expression and cell viability. Polyadenylation of male germ cell mRNAs is unusual, exhibiting increased alternative polyadenylation, decreased AAUAAA polyadenylation signal use, and reduced downstream sequence element dependence. CstF-64, the RNA-binding component of the cleavage stimulation factor (CstF), interacts with pre-mRNAs at sequences downstream of the cleavage site. In mammalian testes, meiotic XY-body formation causes suppression of X-linked CstF-64 expression during pachynema. Consequently, an autosomal paralog, CstF-64 (gene name Cstf2t), is expressed during meiosis and subsequent haploid differentiation. Here we show that targeted disruption of Cstf2t in mice causes aberrant spermatogenesis, specifically disrupting meiotic and postmeiotic development, resulting in male infertility resembling oligoasthenoteratozoospermia. Furthermore, the Cstf2t mutant phenotype displays variable expressivity such that spermatozoa show a broad range of defects. The overall phenotype is consistent with a requirement for CstF-64 in spermatogenesis as indicated by the significant changes in expression of thousands of genes in testes of Cstf2t ؊/؊ mice as measured by microarray. Our results indicate that, although the infertility in Cstf2t ؊/؊ males is due to low sperm count, multiple genes controlling many aspects of germ-cell development depend on CstF-64 for their normal expression. Finally, these transgenic mice provide a model for the study of polyadenylation in an isolated in vivo system and highlight the role of a growing family of testis-expressed autosomal retroposed variants of X-linked genes.spermatogenesis ͉ oligoasthenoteratozoospemia ͉ meiosis ͉ XY body ͉ meiotic sex chromosome inactivation P olyadenylation, the process of mRNA 3Ј end formation, is required for the synthesis, transport, translation, and stability of eukaryotic mRNAs (1, 2). Although polyadenylation is nearly universal, features of polyadenylation are different in mammalian male germ cells than in other tissues: male germ cell mRNAs exhibit increased alternative polyadenylation (3, 4), decreased use of the AAUAAA polyadenylation signal (5, 6), and reduced dependence on downstream sequence elements (DSEs) (7). These differences suggest a modified mechanism for polyadenylation in male germ cells.While examining these differences, we discovered CstF-64 (8), which is a paralog of the 64,000 M r subunit of the cleavage stimulation factor (CstF-64) (9-11). CstF-64 is expressed in nuclei of early spermatogenic cells (5, 12). However, because it is on the X chromosome, CstF-64 expression halts in pachytene spermatocytes because of meiotic sex chromosome inactivation (MSCI) (13). In contrast, CstF-64 expression begins in pachytene spermatocytes and continues in spermatocytes and early spermatids (refs. 5 and 12; summarized in Fig. 1). CstF-64 is the only known CstF-64 homolog expressed during male meiosis, thus making it a candidate to play a critical role in sper...

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Two proteins crosslinked to RNA containing the adenovirus L3 poly(A) site require the AAUAAA sequence for binding.

Cited by 114 publications

References 61 publications

Protein factors in pre-mRNA 3′-end processing

Protein factors in pre-mRNA 3′-end processing

Cleavage Factor II of Saccharomyces cerevisiaeContains Homologues to Subunits of the Mammalian Cleavage/ Polyadenylation Specificity Factor and Exhibits Sequence-specific, ATP-dependent Interaction with Precursor RNA

Loss of polyadenylation protein τCstF-64 causes spermatogenic defects and male infertility

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