TGIF2LY Human

What is TGIF2LY and what are its basic genetic characteristics?

TGIF2LY is a protein-coding gene located on the human Y chromosome, officially known as "TGFB induced factor homeobox 2 like, Y-linked" (synonym: TGIFLY). This gene belongs to the TALE/TGIF homeobox family of transcription factors and is situated within the male-specific region of chromosome Y, in a sequence block that resulted from a large X-to-Y transposition . The gene has an Entrez Gene ID of 90655 and encodes a protein that functions as a transcription factor .

TGIF2LY represents a member of a broader transcription factor family that includes multiple related proteins such as TGIF1, TGIF2, and TGIF2LX . The genomic context of TGIF2LY is particularly noteworthy as it has a homolog on the X chromosome (TGIF2LX), but with significant C-terminal divergence that suggests distinct regulatory functions between these homologs .

How does TGIF2LY structure compare to other TGIF family proteins?

TGIF2LY shares structural similarities with other TGIF family members, particularly in its DNA-binding domain, which is highly conserved. The protein consists of 185 amino acids and contains a characteristic homeodomain that facilitates DNA binding . The three-dimensional structure modeling analyses suggest that TGIF2LY maintains the canonical homeodomain fold found in other TALE homeodomain proteins .

What is the expression pattern of TGIF2LY in normal human tissues?

TGIF2LY demonstrates a restricted expression pattern in normal human tissues, predominantly exhibiting testis-specific expression . This tissue-specific expression profile suggests specialized functions in male reproductive biology. Unlike many Y-chromosome genes with broader expression patterns, TGIF2LY expression is typically limited under normal physiological conditions .

Interestingly, research has demonstrated that TGIF2LY expression may be altered in pathological conditions. For instance, studies have identified upregulated expression in acute myeloid leukemia (AML) cells compared to their healthy counterparts, suggesting potential dysregulation of this gene in hematological malignancies . This differential expression between normal and disease states highlights TGIF2LY as a potential biomarker or therapeutic target in certain contexts.

What DNA sequences does TGIF2LY recognize and how has this been experimentally determined?

TGIF2LY has been shown to recognize and bind specific DNA sequences, consistent with its role as a transcription factor. Research employing electrophoretic mobility shift assays (EMSA) has successfully identified the core nucleotide sequence recognized by TGIF2LY . These studies suggest that TGIF2LY might recognize similar DNA sequences as other TGIF family proteins due to the conservation of its DNA-binding domain .

The experimental determination of TGIF2LY binding sites has involved sophisticated protein-DNA binding assays modified from previously established in vitro genomic selection techniques . These approaches have allowed researchers to isolate genomic DNA fragments that specifically interact with TGIF2LY. Subsequent analysis of these bound fragments has provided insights into potential genes and regulatory elements that might be under TGIF2LY control, suggesting its involvement in cellular differentiation and immune cell activation pathways .

How does TGIF2LY function as a transcription factor compared to its paralog TGIF2LX?

TGIF2LY functions as a DNA-binding transcription factor with potential regulatory effects on gene expression. Subcellular localization studies have confirmed that TGIF2LY is transported to the cell nucleus, consistent with its role in transcriptional regulation . The protein's interaction with specific DNA sequences enables it to influence the expression of target genes involved in various cellular processes.

Comparative analysis suggests that TGIF2LY may act as a competitor or regulator of its X-linked paralog TGIF2LX . This functional relationship is likely facilitated by the significant divergence in their C-terminal regions, which may confer distinct regulatory capabilities or target specificities . Research indicates that despite recognizing similar DNA motifs due to conserved DNA-binding domains, these paralogs might recruit different cofactors or exhibit varying regulatory potentials, potentially allowing for sex-specific gene regulation patterns .

What are the implications of TGIF2LY's Y-chromosome localization for sex-specific gene regulation?

TGIF2LY's location within the male-specific region of chromosome Y has significant implications for sex-specific gene regulation. As a Y-linked transcription factor, TGIF2LY's expression is restricted to males, potentially contributing to male-specific transcriptional programs . This sex-limited expression pattern suggests a role in regulating genes involved in male development, reproduction, or other sexually dimorphic traits.

The evolutionary relationship between TGIF2LY and its X-linked homolog TGIF2LX represents a classic example of sex chromosome divergence following an ancestral transposition event . The functional divergence between these paralogs might contribute to differences in gene regulation between males and females. Additionally, the unique regulatory properties of TGIF2LY might influence sex-specific disease susceptibilities or therapeutic responses, particularly in conditions where TGIF family proteins play significant roles .

What cell models and experimental systems are optimal for studying TGIF2LY function?

Research on TGIF2LY has employed various cell models, with THP-1 cells emerging as a particularly useful system . THP-1, a human monocytic cell line derived from an acute monocytic leukemia patient, has been validated to express TGIF2LY protein and thus serves as an accessible model for investigating TGIF2LY function . This cell line allows researchers to study TGIF2LY in a controlled environment that reflects some aspects of its pathological expression in leukemia.

For investigating the normal physiological functions of TGIF2LY, primary testicular cells or testis-derived cell lines might provide more relevant models, given the gene's predominant expression in testis under normal conditions . Additionally, comparative studies between male and female cells (particularly those with TGIF2LX expression) can yield insights into the sex-specific regulatory functions of these paralogs. Experimental systems utilizing CRISPR-Cas9 gene editing to modify TGIF2LY expression or structure also offer powerful approaches for functional characterization .

What techniques are most effective for detecting and quantifying TGIF2LY expression?

Real-time PCR with low-density arrays has proven effective for detecting and quantifying TGIF2LY expression at the transcript level . When designing primers and probes for TGIF2LY detection, researchers must carefully consider the high sequence similarity with its X-linked homolog to ensure specificity. TaqMan probes with appropriate controls have been successfully employed to distinguish TGIF2LY expression from related genes .

For protein-level detection, western blotting using specific antibodies against TGIF2LY has been utilized, particularly in experimental contexts like THP-1 cells . Recombinant TGIF2LY protein expressed in Escherichia coli systems has served as useful standards for such analyses . Immunohistochemistry and immunofluorescence approaches can provide spatial information about TGIF2LY expression in tissues and subcellular localization within cells, complementing quantitative expression data with valuable contextual information .

How can researchers effectively study TGIF2LY's DNA-binding properties and target genes?

Investigating TGIF2LY's DNA-binding properties requires specialized techniques adapted for transcription factor research. Electrophoretic mobility shift assays (EMSA) have been successfully employed to identify the core nucleotide sequences recognized by TGIF2LY . These assays involve incubating purified TGIF2LY protein with labeled DNA fragments and analyzing the resulting protein-DNA complexes through gel electrophoresis.

For genome-wide identification of TGIF2LY binding sites, modified protein-DNA binding assays derived from in vitro genomic selection techniques have proven valuable . These approaches can be complemented with chromatin immunoprecipitation followed by sequencing (ChIP-seq) in appropriate cellular models expressing TGIF2LY. Subsequent bioinformatic analysis of identified binding regions can reveal enriched motifs and potential target genes, providing insights into the biological pathways regulated by TGIF2LY .

What is known about TGIF2LY's role in hematological malignancies, particularly AML?

Research has identified altered expression of TGIF2LY in acute myeloid leukemia (AML) cells compared to healthy counterparts, suggesting a potential role in leukemia pathogenesis . Specifically, TGIF2LY has been found to be up-regulated in certain AML subtypes, particularly M4 and M5, which are characterized by high CD14 expression on leukemic cell surfaces . This abnormal expression pattern is notable considering TGIF2LY's normally restricted expression profile.

The mechanistic implications of TGIF2LY upregulation in AML remain under investigation. Given its transcription factor function, increased TGIF2LY expression might dysregulate genes involved in hematopoietic differentiation or cellular proliferation . Analysis of genomic DNA fragments bound by TGIF2LY suggests a potential role in the differentiation and activation of antigen-presenting cells, which might be relevant to leukemia biology . These findings position TGIF2LY as a potential biomarker or therapeutic target in certain AML contexts, particularly in male patients.

How does TGIF2LY's expression pattern differ between normal and pathological states?

TGIF2LY exhibits markedly different expression patterns between normal and pathological states. Under normal physiological conditions, TGIF2LY expression is predominantly restricted to testicular tissue, reflecting its specialized functions in male reproductive biology . This tissue-specific expression is tightly regulated and maintained under normal developmental and physiological conditions.

In contrast, pathological states such as acute myeloid leukemia (AML) have been associated with aberrant TGIF2LY expression . Studies comparing AML cells with healthy counterparts have identified significant upregulation of TGIF2LY in leukemic cells, particularly in specific AML subtypes . This pathological expression pattern suggests potential dysregulation of TGIF2LY's normal transcriptional control mechanisms in cancer cells, which might contribute to altered gene expression programs that support malignant phenotypes.

What potential therapeutic applications might target TGIF2LY in disease contexts?

TGIF2LY's restricted normal expression pattern combined with its aberrant expression in certain malignancies presents intriguing therapeutic possibilities. In hematological malignancies like AML where TGIF2LY is upregulated, targeted inhibition of TGIF2LY function might represent a novel therapeutic approach . Such strategies could include small molecule inhibitors that disrupt TGIF2LY's DNA-binding capabilities or protein-protein interactions essential for its transcriptional activities.

What are the key considerations when working with recombinant TGIF2LY protein?

Working with recombinant TGIF2LY protein requires attention to several technical considerations. Commercially available recombinant Human TGIF2LY protein, such as the full-length protein (amino acids 1-185) expressed in Escherichia coli, provides a valuable research tool with >95% purity suitable for various applications including SDS-PAGE and mass spectrometry . When selecting or producing recombinant TGIF2LY, researchers should consider:

• Expression system compatibility: E. coli-expressed TGIF2LY lacks eukaryotic post-translational modifications, which may be relevant for certain functional studies.

• Purification tags: Common tags like polyhistidine (His-tag) facilitate purification but may influence protein behavior in certain assays.

• Proper folding verification: Ensuring the recombinant protein adopts its native conformation, particularly within the critical DNA-binding homeodomain.

• Storage conditions: Typically requiring -80°C storage with minimal freeze-thaw cycles to preserve activity.

What methodological approaches have yielded insights into TGIF2LY's function as a transcription factor?

Multiple methodological approaches have contributed to our understanding of TGIF2LY's transcription factor function. Subcellular localization studies demonstrating nuclear transport of TGIF2LY provide foundational evidence supporting its role in transcriptional regulation . Amino acid sequence analysis and three-dimensional structure modeling have further reinforced this functional classification by identifying conserved DNA-binding domains characteristic of transcription factors .

Protein-DNA interaction studies have been particularly informative. Modified protein-DNA binding assays derived from in vitro genomic selection techniques have allowed identification of genomic DNA regions bound by TGIF2LY . Electrophoretic mobility shift assays (EMSA) have subsequently facilitated the identification of core nucleotide sequences specifically recognized by TGIF2LY . Together, these approaches have not only confirmed TGIF2LY's capacity to bind DNA but have also provided insights into its potential target genes and regulatory networks.

What bioinformatic approaches are useful for predicting TGIF2LY target genes and biological pathways?

Bioinformatic approaches offer powerful means to predict TGIF2LY target genes and associated biological pathways. Analysis of experimentally identified TGIF2LY-bound genomic DNA fragments suggests potential roles in differentiation and activation of antigen-presenting cells . Several bioinformatic strategies can extend these insights:

• Motif analysis of confirmed binding sites to generate position weight matrices for genome-wide prediction of potential binding sites.

• Integration with gene expression data from contexts where TGIF2LY is manipulated (overexpression, knockdown) to correlate binding with transcriptional outcomes.

• Gene ontology and pathway enrichment analysis of predicted targets to identify biological processes potentially regulated by TGIF2LY.

• Comparative genomics approaches examining conservation of binding sites across species to prioritize functionally significant targets.

• Network analysis integrating TGIF2LY with known protein interactors and co-regulators to construct comprehensive regulatory networks.

These computational approaches complement experimental findings and can guide further investigation into TGIF2LY's biological functions and disease relevance.