Crystal structure of the DNA-binding domain of Myelin-gene Regulatory Factor

Zhen, Xiangkai; Li, Bowen; Hu, Fang; Yan, Shufeng; Meloni, Gabriele; Li, Hui‐Liang; Shi, Ning

doi:10.1038/s41598-017-03768-9

Cited by 13 publications

(16 citation statements)

References 32 publications

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“…For the control set, we were able to detect the N-terminal fragments of Flag-Myrf, Myc-Myrf, and HA-Myrf in the MB fraction ( Fig. 5D), consistent with its homo-trimerization 21,24,25 . The specificity of the second Myc immunoprecipitation step was confirmed by the lack of immunoblot signals in the MB fraction for the Set 3 samples.…”

Section: Scientific Reports |supporting

confidence: 57%

Functional mechanism and pathogenic potential of MYRF ICA domain mutations implicated in birth defects

Fan

Sharif

et al. 2020

Sci Rep

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Myrf is a membrane-bound transcription factor that plays a key role in various biological processes. the intramolecular chaperone Auto-processing (icA) domain of Myrf forms a homo-trimer, which carries out the auto-cleavage of Myrf. the icA homo-trimer-mediated auto-cleavage of Myrf is a prerequisite for its transcription factor function in the nucleus. Recent exome sequencing studies have implicated two MYRF ICA domain mutations (V679A and R695H) in a novel syndromic form of birth defects. It remains unknown whether and how the two mutations impact the transcription factor function of Myrf and, more importantly, how they are pathogenic for congenital anomalies. Here, we show that V679A and R695H cripple the ICA domain, blocking the auto-cleavage of Myrf. Consequently, Myrf-V679A and Myrf-R695H do not exhibit any transcriptional activity. Molecular modeling suggests that V679A and R695H abrogate the auto-cleavage function of the ICA homo-trimer by destabilizing its homo-trimeric assembly. We also found that the ICA homo-trimer can tolerate one copy of Myrf-V679A or Myrf-R695H for its auto-cleavage function, indicating that V679A and R695H are not dominant negatives. Thus, if V679A and R695H in a heterozygous state caused birth defects, it would be via haploinsufficiency of MYRF.Myrf (myelin regulatory factor, previously known as C11orf9 [the human gene] and Mrf [the mouse gene]) is a pleiotropic membrane-bound transcription factor 1-7 . Throughout this paper, MYRF and Myrf refer to the human and mouse genes, respectively. In the murine central nervous system (CNS), Myrf is specifically expressed by oligodendrocytes (OLs) 1,8 . Conditional knockout of Myrf in OL lineage cells led to widespread dysmyelination and severe neurological deficits 1 . Myrf is also indispensable for the life-long maintenance, plasticity, and regeneration of CNS myelin 9-11 . These studies left the impression that Myrf is a "myelin" transcription factor, and hence the name Myrf. We now know that MYRF is also expressed in other tissues such as stomach, lung, heart, ovary, eye, and developing gonads [12][13][14][15] . Consistently, MYRF coding variants have been implicated in both myelin and non-myelin diseases [13][14][15][16][17][18][19] , and whole-body Myrf knockout led to embryonic lethality in mouse independent of OL development 1 . In keeping with myelin-independent functions of Myrf, Myrf orthologs are found and play an important role in organisms without myelin 2,3,6,7 .Myrf is generated as a type-II membrane protein in the endoplasmic reticulum (ER) 4-6 . The Intramolecular Chaperone Auto-processing (ICA) domain is pivotal to the auto-cleavage of Myrf 4,5,20 . The ICA domain forms a homo-trimer, which as an intramolecular chaperone helps Myrf to form a homo-trimer in the ER membrane by chaperoning the formation of a triple β-helix (the intramolecular chaperone function, Fig. 1) 4,5,20-23 . As soon as the triple β-helix is completed, the ICA homo-trimer performs the auto-cleavage reactions that cleave the three P586-S587 peptide bon...

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Section: Scientific Reports |supporting

confidence: 57%

Functional mechanism and pathogenic potential of MYRF ICA domain mutations implicated in birth defects

Fan

Sharif

et al. 2020

Sci Rep

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“…In contrast, the nuclear entry step seems to be under regulation for the worm and slime mold orthologs of Myrf 4 , 8 , 9 . A recent structural study confirmed the homo-trimerization status for Myrf N-terminal fragments 16 , and we found that homo-trimerization imparts DNA-binding specificity to Myrf N-terminal fragment 15 , which provides a rational explanation for the lethal phenotype of the let-25 allele of C . elegans 8 .…”

Section: Introductionsupporting

confidence: 78%

Elucidating the transactivation domain of the pleiotropic transcription factor Myrf

Choi

Fan

Kim

et al. 2018

Sci Rep

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Myrf is a newly discovered membrane-bound transcription factor that plays an essential role in as diverse organisms as human, worm, and slime mold. Myrf is generated as a type-II membrane protein in the endoplasmic reticulum (ER). It forms homo-oligomers to undergo auto-cleavage that releases Myrf N-terminal fragment from the ER membrane as a homo-trimer. The homo-trimer of Myrf N-terminal fragments enters the nucleus and binds the Myrf motif to activate transcription. Despite its prominent role as a transcriptional activator, little is known about the transactivation domain of Myrf. Here, we report that the N-terminal-most (NTM) domain of Myrf is required for transcriptional activity and, when fused to a Gal4 fragment, enables it to activate transcription. The transactivation function of the NTM domain did not require homo-trimerization. We also discovered that the NTM domain can be sumoylated at three lysine residues (K123, K208, and K276), with K276 serving as the main acceptor. K276 sumoylation repressed the transactivation function of the NTM domain without affecting the stability or nuclear localization of Myrf N-terminal fragment. In sum, this study identifies the NTM domain as the transactivation domain of Myrf and the potential regulatory impact of its K276 sumoylation.

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“…First, in this study, we examined only SNPs located within the NOS2 gene region. Early research has shown that regions 20 kb up- and downstream of genes might contain some important regulatory elements and that variants within these regions could significantly affect gene expression levels [ 26 – 28 ]. Given of the limitations of single SNP analyses [ 29 – 35 ], for future replication studies, it is necessary to conduct haplotype analyses to provide further statistical evidence for our findings, and it is also important to consider those regions and investigate their potential contributions to the risk of non-union in the fracture healing process thoroughly.…”

Section: Discussionmentioning

confidence: 99%

Genetic polymorphisms of NOS2 and predisposition to fracture non-union: A case control study based on Han Chinese population

et al. 2018

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A non-union, especially atrophic non-unions, is a permanent failure of healing following a fracture and can be difficult to treat. Approximately 5–10% of fractures will result in a non-union during the healing process. non-unions can be classified into two types: atrophic non-union which is often due to impaired bone healing with a potential biological mechanism, and hypertrophic non-union which is due to inadequate fixation after fracture. Genetic variations also play an important role in the fracture healing response. Previous studies based on animal models have indicated that NOS2 might be greatly involved in the bone fracture healing process. In this case-control study, 346 nonunion patients were compared to 883 patients with normal fracture healing to investigate the potential genetic association between NOS2 and the fracture healing process using study subjects of Chinese Han ancestry. Twenty-seven single nucleotide polymorphisms (SNPs) covering NOS2 were genotyped in our study subjects and analyzed. In addition to the single marker-based analysis, we performed a gene-by-environment analysis to examine the potential interactions between genetic polymorphisms and some environmental factors. SNP rs2297514 showed significant association with the fracture healing process after adjusting for age and gender (OR = 1.38, P = 0.0005). Our results indicated that the T allele of rs2297514 significantly increased the risk of a non-union during the fracture healing process by 38% compared to the C allele. Further stratification analyses conducted for this SNP using data from subgroups classified by different sites of fracture indicated that significance could only be observed in the tibial diaphysis subgroup (N = 428, OR = 1.77, P = 0.0007) but not other groups including femur diaphysis, humeral shaft, ulnar shaft, and femur neck. Gene-by-environment interaction analyses of the three environmental factors showed no significant results. In this study, rs2297514 was significantly associated with the non-union status of fracture healing using a large Chinese population-based study sample. Our findings replicated those of a previous preliminary study and offered strong evidence linking NOS2 and fracture healing.

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Crystal structure of the DNA-binding domain of Myelin-gene Regulatory Factor

Cited by 13 publications

References 32 publications

Functional mechanism and pathogenic potential of MYRF ICA domain mutations implicated in birth defects

Functional mechanism and pathogenic potential of MYRF ICA domain mutations implicated in birth defects

Elucidating the transactivation domain of the pleiotropic transcription factor Myrf

Genetic polymorphisms of NOS2 and predisposition to fracture non-union: A case control study based on Han Chinese population

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