A Conserved Interaction That Is Essential for the Biogenesis of Histone Locus Bodies

Yang, Xiao Cui; Sabath, Ivan; Kunduru, Lalitha; Wijnen, André J. van; Marzluff, William F.; Domiński, Zbigniew

doi:10.1074/jbc.m114.616466

Cited by 38 publications

(70 citation statements)

References 71 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We recently demonstrated that the C-terminal domain of FLASH directly interacts with the last 25 amino acids of NPAT [24]. Interestingly, a homologous SANT/Myb-like domain capable of interacting with the same region of NPAT also exists at the end of the unrelated vertebrate protein Yin Yang 1-associated protein-related protein (YARP).…”

Section: Introductionmentioning

confidence: 99%

Mapping the Interaction Network of Key Proteins Involved in Histone mRNA Generation: A Hydrogen/Deuterium Exchange Study

Skrajna

Yang

Tarnowski

et al. 2016

Journal of Molecular Biology

Self Cite

View full text Add to dashboard Cite

Histone pre-mRNAs are cleaved at the 3′ end by a complex that contains U7 snRNP, the FLICE-Associated Huge protein (FLASH) and Histone pre-mRNA Cleavage Complex (HCC) consisting of several polyadenylation factors. Within the complex, the N-terminus of FLASH interacts with the N-terminus of the U7 snRNP protein Lsm11 and together they recruit the HCC. FLASH through its distant C-terminus independently interacts with the C-terminal SANT/Myb-like domain of Nuclear Protein, Ataxia-Telangiectasia locus (NPAT), a transcriptional co-activator required for expression of histone genes in S-phase. To gain structural information on these interactions, we used mass spectrometry to monitor hydrogen/deuterium (H/D) exchange in various regions of FLASH, Lsm11 and NPAT alone or in the presence of their respective binding partners. Our results indicate that the FLASH-interacting domain in Lsm11 is highly dynamic, while the more downstream region required for recruiting the HCC exchanges deuterium slowly and likely folds into a stable structure. In FLASH, a stable structure is adopted by the domain that interacts with Lsm11 and this domain is further stabilized by binding Lsm11. Notably, both H/D exchange experiments and in vitro binding assays demonstrate that Lsm11, in addition to interacting with the N-terminal region of FLASH, also contacts the C-terminal SANT/Myb-like domain of FLASH, the same region that binds NPAT. However, while NPAT stabilizes this domain, Lsm11 causes its partial relaxation. These competing reactions may play a role in regulating histone gene expression in vivo.

show abstract

Section: Introductionmentioning

confidence: 99%

Mapping the Interaction Network of Key Proteins Involved in Histone mRNA Generation: A Hydrogen/Deuterium Exchange Study

Skrajna

Yang

Tarnowski

et al. 2016

Journal of Molecular Biology

Self Cite

View full text Add to dashboard Cite

show abstract

“…with »500,000 for U1 snRNA in mammals) and very small amounts of FLASH in Drosophila embryos and larvae. 72 FLASH is recruited to the HLB by an interaction between the C-terminus of FLASH and the C-terminus of NPAT/Mxc 29,44,72,73 . A hypomorphic allele of mxc (mxc G46 ) that encodes a truncated form of Mxc lacking the last 195 amino acids results in failure of full length, wild type FLASH protein and U7 snRNP to concentrate in the HLB, although an HLB containing the truncated Mxc protein forms.…”

Section: Hlb Functionmentioning

confidence: 99%

“…42 A second example is muscle wasted (Mute), discovered by the Chia lab in a screen for factors required for muscle development and shown to concentrate in the HLB, 43 as does its mammalian ortholog YARP. 44 Mute may function as a negative regulator of histone gene expression. 43 Dominski and colleagues also showed that cleavage of histone pre-mRNA is catalyzed by CPSF-73, the same protein that catalyzes cleavage of pre-mRNAs encoding poly(A) mRNAs, demonstrating a role for polyadenylation factors in histone pre-mRNA processing, 45 including Symplekin 46 and CPSF100.…”

Section: An Expanding List Of Hlb Componentsmentioning

confidence: 99%

“…Mammalian YARP, the Drosophila homolog of Mute, and FLASH each interact with the C-terminus of NPAT using a Myb-like domain, and this domain from either protein can localize a heterologous protein to the HLB. 44 The C-terminus of NPAT can bind either YARP or FLASH but not both at once. Detailed in vitro binding studies demonstrated that the C-terminal Myb-like domain of FLASH also binds to Lsm11 adjacent to the binding site on Lsm11 for the N-terminus of FLASH, and that this interaction prevents FLASH binding to NPAT.…”

Section: Knockdown Of Y3mentioning

confidence: 99%

“…Taking advantage of our knowledge of the molecular interactions of HLB components, 44,70,72,73,[99][100][101] we have performed genetic studies in Drosophila using mutant proteins that are either processing defective or unable to localize to the HLB. Unlike other metazoan genes, where transcription can continue well 3 0 of the polyadenylation site, transcription termination is tightly coupled to histone mRNA processing in both Drosophila and mammals.…”

Section: Hlb Functionmentioning

confidence: 99%

See 2 more Smart Citations

Coordinating cell cycle-regulated histone gene expression through assembly and function of the Histone Locus Body

2017

Self Cite

View full text Add to dashboard Cite

Metazoan replication-dependent (RD) histone genes encode the only known cellular mRNAs that are not polyadenylated. These mRNAs end instead in a conserved stem-loop, which is formed by an endonucleolytic cleavage of the pre-mRNA. The genes for all 5 histone proteins are clustered in all metazoans and coordinately regulated with high levels of expression during S phase. Production of histone mRNAs occurs in a nuclear body called the Histone Locus Body (HLB), a subdomain of the nucleus defined by a concentration of factors necessary for histone gene transcription and premRNA processing. These factors include the scaffolding protein NPAT, essential for histone gene transcription, and FLASH and U7 snRNP, both essential for histone pre-mRNA processing. Histone gene expression is activated by Cyclin E/Cdk2-mediated phosphorylation of NPAT at the G1-S transition. The concentration of factors within the HLB couples transcription with pre-mRNA processing, enhancing the efficiency of histone mRNA biosynthesis. KEYWORDSCell cycle; Drosophila; histone genes; mRNA processing; nuclear bodyProper utilization of genetic information in eukaryotes requires extensive three-dimensional organization of the genome and its associated proteins within the nucleus. The basic unit of genome organization is the nucleosome, an octamer of two molecules of each of the four core histone proteins (H2A, H2B, H3 and H4) that packages »147 base pairs of DNA. In mammalian cells, approximately 4£10 8 molcules of each core histone must be synthesized during S phase of the cell cycle to package the newly replicated DNA. Defects in this process result in genome instability that can contribute to human pathologies like cancer. Histone protein synthesis results from a large increase in production at the beginning of S phase of equal amounts of mRNAs encoding the four core nucleosomal histones, as well as histone H1. This highly coordinated gene expression event is performed by a set of transcription and pre-mRNA processing factors unique to replication dependent (RD) histone mRNA biosynthesis that assemble into a nuclear body tightly associated with RD histone genes called the histone locus body (HLB). HLB formation involves a combination of ordered and stochastic assembly steps, and cell cycle regulated changes in the HLB occur to activate histone gene expression during S phase. Here we describe the history of how the HLB was discovered followed by a discussion of HLB assembly mechanism and how the HLB functions in RD histone mRNA biosynthesis.

show abstract

Intranuclear and higher‐order chromatin organization of the major histone gene cluster in breast cancer

Fritz

Ghule

Boyd

et al. 2017

Journal Cellular Physiology

Self Cite

View full text Add to dashboard Cite

Alterations in nuclear morphology are common in cancer progression. However, the degree to which gross morphological abnormalities translate into compromised higher-order chromatin organization is poorly understood. To explore the functional links between gene expression and chromatin structure in breast cancer, we performed RNA-seq gene expression analysis on the basal breast cancer progression model based on human MCF10A cells. Positional gene enrichment identified the major histone gene cluster at chromosome 6p22 as one of the most significantly upregulated (and not amplified) clusters of genes from the normal-like MCF10A to premalignant MCF10AT1 and metastatic MCF10CA1a cells. This cluster is subdivided into three sub-clusters of histone genes that are organized into hierarchical topologically associating domains (TADs). Interestingly, the sub-clusters of histone genes are located at TAD boundaries and interact more frequently with each other than the regions in-between them, suggesting that the histone sub-clusters form an active chromatin hub. The anchor sites of loops within this hub are occupied by CTCF, a known chromatin organizer. These histone genes are transcribed and processed at a specific sub-nuclear microenvironment termed the major histone locus body (HLB). While the overall chromatin structure of the major HLB is maintained across breast cancer progression, we detected alterations in its structure that may relate to gene expression. Importantly, breast tumor specimens also exhibit a coordinate pattern of upregulation across the major histone gene cluster. Our results provide a novel insight into the connection between the higher-order chromatin organization of the major HLB and its regulation during breast cancer progression.

show abstract

A Conserved Interaction That Is Essential for the Biogenesis of Histone Locus Bodies

Cited by 38 publications

References 71 publications

Mapping the Interaction Network of Key Proteins Involved in Histone mRNA Generation: A Hydrogen/Deuterium Exchange Study

Mapping the Interaction Network of Key Proteins Involved in Histone mRNA Generation: A Hydrogen/Deuterium Exchange Study

Coordinating cell cycle-regulated histone gene expression through assembly and function of the Histone Locus Body

Intranuclear and higher‐order chromatin organization of the major histone gene cluster in breast cancer

Contact Info

Product

Resources

About