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

Skrajna, Aleksandra; Yang, Xiao Cui; Tarnowski, Krzysztof; Fituch, Kinga; Marzluff, William F.; Domiński, Zbigniew; Dadlez, Michał

doi:10.1016/j.jmb.2016.01.031

Cited by 9 publications

(12 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This α-helical fold might also extend to residues 53–70, encompassing the LDLY motif, but this region is unlikely to contribute to the dimerization interface. Our data are consistent with recent H/D exchange studies [ 33 ], which showed that residues 75–136 underwent slow H/D exchange, indicative of extensive secondary structure in this region. Residues 58–62 exchanged significantly faster than the 75–136 region but slower than the directly surrounding sequences, suggesting the presence of a more dynamic secondary structure in the vicinity of the LDLY motif [ 33 ].…”

Section: Discussionsupporting

confidence: 93%

“…That Lsm11 interacts with a FLASH dimer is also consistent with the data from H/D exchange experiments. While the region between amino acids 100–120 showed the slowest H/D exchange within the entire FLASH NTD (which we show here can form a dimer), this region underwent slower H/D exchange in the presence of Lsm11 and the reduced rate of exchange extended to amino acid 130 in FLASH [ 33 ]. Since H/D exchange occurs when hydrogen bonds are temporarily destabilized, this region of FLASH (residues 100–130) is in a more stable structure in the heterotrimer than in the homodimer.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

The N-terminal domains of FLASH and Lsm11 form a 2:1 heterotrimer for histone pre-mRNA 3’-end processing

Aik

Lin²,

Tan

et al. 2017

PLoS ONE

Self Cite

View full text Add to dashboard Cite

Unlike canonical pre-mRNAs, animal replication-dependent histone pre-mRNAs lack introns and are processed at the 3’-end by a mechanism distinct from cleavage and polyadenylation. They have a 3’ stem loop and histone downstream element (HDE) that are recognized by stem-loop binding protein (SLBP) and U7 snRNP, respectively. The N-terminal domain (NTD) of Lsm11, a component of U7 snRNP, interacts with FLASH NTD and these two proteins recruit the histone cleavage complex containing the CPSF-73 endonuclease for the cleavage reaction. Here, we determined crystal structures of FLASH NTD and found that it forms a coiled-coil dimer. Using solution light scattering, we characterized the stoichiometry of the FLASH NTD-Lsm11 NTD complex and found that it is a 2:1 heterotrimer, which is supported by observations from analytical ultracentrifugation and crosslinking.

show abstract

Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

The N-terminal domains of FLASH and Lsm11 form a 2:1 heterotrimer for histone pre-mRNA 3’-end processing

Aik

Lin²,

Tan

et al. 2017

PLoS ONE

Self Cite

View full text Add to dashboard Cite

show abstract

“…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%

“…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. 99 Thus, it is tempting to speculate that reorganization of the Lsm11-FLASH-NPAT interactions in the HLB may be part of U7 snRNP activation and the onset of RD histone premRNA processing during S phase. This idea also suggests that different pools of molecules may exist within the HLB.…”

Section: Knockdown Of Y3mentioning

confidence: 99%

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

“…1A). It is unclear whether the holo-U7 snRNP contains all subunits necessary for processing and whether its assembly is regulated during the cell cycle, with FLASH binding Lsm11 during the G1/S transition, as suggested by Skrajna et al (2016).…”

Section: Introductionmentioning

confidence: 99%

U7 snRNP is recruited to histone pre-mRNA in a FLASH-dependent manner by two separate regions of the stem–loop binding protein

Skrajna

Yang

Bucholc

et al. 2017

RNA

Self Cite

View full text Add to dashboard Cite

Cleavage of histone pre-mRNAs at the 3' end requires stem-loop binding protein (SLBP) and U7 snRNP that consists of U7 snRNA and a unique Sm ring containing two U7-specific proteins: Lsm10 and Lsm11. Lsm11 interacts with FLASH and together they bring a subset of polyadenylation factors to U7 snRNP, including the CPSF73 endonuclease that cleaves histone pre-mRNA. SLBP binds to a conserved stem-loop structure upstream of the cleavage site and acts by promoting an interaction between the U7 snRNP and a sequence element located downstream from the cleavage site. We show that both human and SLBPs stabilize U7 snRNP on histone pre-mRNA via two regions that are not directly involved in recognizing the stem-loop structure: helix B of the RNA binding domain and the C-terminal region that follows the RNA binding domain. Stabilization of U7 snRNP binding to histone pre-mRNA by SLBP requires FLASH but not the polyadenylation factors. Thus, FLASH plays two roles in 3' end processing of histone pre-mRNAs: It interacts with Lsm11 to form a docking platform for the polyadenylation factors, and it cooperates with SLBP to recruit U7 snRNP to histone pre-mRNA.

show abstract

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

Cited by 9 publications

References 37 publications

The N-terminal domains of FLASH and Lsm11 form a 2:1 heterotrimer for histone pre-mRNA 3’-end processing

The N-terminal domains of FLASH and Lsm11 form a 2:1 heterotrimer for histone pre-mRNA 3’-end processing

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

U7 snRNP is recruited to histone pre-mRNA in a FLASH-dependent manner by two separate regions of the stem–loop binding protein

Contact Info

Product

Resources

About