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Biology subjects

Nemcko, F.

Publications and source records attributed to Nemcko, F..

6 recordsLinked to original sources

BioReason-Pro: Advancing Protein Function Prediction with Multimodal Biological Reasoning

Protein function annotation is fundamental to understanding biological mechanisms, designing therapeutics, and advancing biomedical research. Current computational methods either rely on shallow sequence similarity or treat function prediction as isolated classification tasks, failing to capture the integrative reasoning across sequence, structure, domains, and interactions that expert biologists perform to infer function. We introduce BioReason-Pro, the first multimodal reasoning large language model (LLM) for protein function prediction that integrates protein embeddings with biological context to generate structured reasoning traces. A key input into BioReason-Pro is the set of GO term predictions made by GO-GPT, our autoregressive transformer that captures hierarchical and cross-aspect dependencies of GO terms. BioReason-Pro is trained via supervised fine-tuning on synthetic reasoning traces generated by GPT-5 for over 130K proteins and further optimized through reinforcement learning. It achieves 73.6% Fmax on GO term prediction and an LLM judge score of 8/10 on functional summaries, substantially outperforming previous methods. Evaluations with human protein experts show that BioReason-Pro annotations are preferred over ground truth UniProt annotations in 79% of cases. Remarkably, BioReason-Pro predicted a novel interaction partner for the renal cancer biomarker RCDG1, which we confirmed in the lab by co-immunoprecipitation. In other binding-partner predictions, its per-residue attention localized to the exact contact residues resolved in cryo-EM structures. Together, GO-GPT and BioReason-Pro establish a framework for protein function prediction that combines precise ontology modeling with interpretable biological reasoning.

molecular biology↗

Activator-promoter compatibility in mammals: a CpG-Island-specific co-activator directly bridges transcription factors to TFIID

Transcription from CpG island (CGI) promoters controls the expression of two-thirds of mammalian genes, yet despite their prevalence, it remains unknown whether CGI-specific co-activators with intrinsic specificity (i.e. compatibility) for these promoters exist or by what mechanisms they might function. Here, we perform proteome-wide functional screens to identify more than fifty transcriptional activators that are intrinsically specific to CGI promoters, establishing that promoter-class-specific activators are a widespread feature of mammalian gene regulation. Among these, we identify Host Cell Factor 1 (Hcfc1) as the founding member of CGI-specific co-activators. Hcfc1 is essential for the expression of thousands of CGI-promoter-driven genes and acquires CGI-specificity through a two-step mechanism: CGI-associated transcription factors recruit Hcfc1 through its Kelch domain, and Hcfc1 in turn directly engages the general transcription factor TFIID through a dedicated activation domain. The Hcfc1-TFIID interaction overcomes a key rate-limiting step for CGI promoter initiation, TFIID recruitment, thereby directly enabling transcription. Hcfc1 thus functions as a promoter-class-specific bridge between CGI-bound transcription factors and the general transcription machinery, analogous to Mediator but with intrinsic promoter specificity. Together, we uncover a dedicated activation pathway for CGI promoters, reveal a fundamental mechanistic difference in transcription activation between promoter classes in mammals, and establish co-activator-promoter compatibility as a core principle in mammalian gene regulation.

molecular biology↗

RNA decay via the nuclear exosome is essential for piwi-mediated transposon silencing

Nuclear Argonaute proteins safeguard genome integrity by directing transcriptional silencing and heterochromatin formation at transposon loci. Yet it remains unclear how Argonautes enforce robust repression while relying on target transcription for their own recruitment. Here we show that transposon silencing by the Drosophila nuclear Piwi-piRNA pathway requires degradation of target RNA by the nuclear exosome. Using proximity proteomics at endogenous Piwi target sites, we identify two previously uncharacterized paralogs, TEsup-1 and TEsup-2, as essential cofactors for Piwi-mediated silencing. TEsup proteins act in part by engaging nuclear exosome adaptor complexes at piRNA-targeted transcripts through a domain that recognizes proline-rich peptides. Disruption of the Piwi-TEsup-exosome axis leads to accumulation and nuclear export of piRNA-targeted transposon RNAs. Notably, the P-element--which evades heterochromatin-based repression--is silenced primarily through this RNA-decay pathway. Thus, the nuclear piRNA pathway couples target recognition to RNA degradation, reconciling small RNA-guided heterochromatin formation with ongoing transcription at target loci.

molecular biology↗

Proteome-scale tagging and functional screening in mammalian cells by ORFtag

Determining protein function in a systematic manner is a key goal of modern biology, but remains challenging with current approaches. Here, we present ORFtag, a versatile, cost-effective and highly efficient method for the massively-parallel tagging and functional interrogation of proteins at proteome scale. Using mouse embryonic stem cells, we showcase ORFtags utility through screens for transcriptional activators, repressors and post-transcriptional regulators. Each screen finds known and novel regulators, including long ORFs not accessible to other methods, revealing that Zfp574 is a highly selective transcriptional activator and that oncogenic fusions frequently function as transactivators.

systems biology↗

Functionally distinct promoter classes initiate transcription via different mechanisms reflected in focused versus dispersed initiation patterns

Recruitment of RNA polymerase II (Pol II) to promoter regions is essential for transcription. Despite conflicting evidence, the Pol II Pre-Initiation Complex (PIC) is often thought to be of uniform composition and assemble at all promoters via an identical mechanism. Here, we show using Drosophila melanogaster S2 cells as a model that promoter classes with distinct functions and initiation patterns function via PICs that display different compositions and dependencies: developmental promoter DNA readily associates with the canonical Pol II PIC, whereas housekeeping promoter DNA does not and instead recruit different factors such as DREF. Consistently, TBP and DREF are required by distinct sets of promoters, and TBP and its paralog TRF2 function at different promoter types, partly exclusively and partly redundantly. In contrast, TFIIA is required for transcription from all promoters, and we identify factors that can recruit and/or stabilize TFIIA at housekeeping promoters and activate transcription. We show that promoter activation by these factors is sufficient to induce the dispersed transcription initiation patterns characteristic of housekeeping promoters. Thus, different promoter classes direct distinct mechanisms of transcription initiation, which relate to different focused versus dispersed initiation patterns.

molecular biology↗

Identification and characterization of repressive domains in Drosophila transcription factors

All multicellular life relies on differential gene expression, determined by regulatory DNA elements and DNA-binding transcription factors that mediate activation and repression via cofactor recruitment. While activators have been extensively characterized, repressors are less well studied and their repressive domains (RDs) are typically unknown, as are the RDs properties and the co-repressors (CoRs) they recruit. Here, we develop the high-throughput next-generation-sequencing-based method Repressive-Domain (RD)-seq to systematically identify RDs in complex libraries. Screening more than 200,000 fragments covering the coding sequences of all transcription-related proteins in Drosophila melanogaster, we identify 195 RDs in known repressors and in proteins not previously associated with repression. Many RDs contain recurrent short peptide motifs that are required for RD function, as demonstrated by motif mutagenesis, and are conserved between fly and human. Moreover, we show that RDs which contain one of five distinct repressive motifs interact with and depend on different CoRs, including Groucho, CtBP, Sin3A or Smrter. Overall, our work constitutes an invaluable resource and advances our understanding of repressors, their sequences, and the functional impact of sequence-altering mutations.

molecular biology↗