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Comparisons of clinical isolate genomes previously identified a gene,\n                      <jats:italic toggle=\"yes\">msf<\/jats:italic>\n                      , that contained Sel1-like repeats that were associated with clinical disease. Mutant analysis had further found that\n                      <jats:italic toggle=\"yes\">msf<\/jats:italic>\n                      improved survival in macrophages and increased systemic infection in an animal model. However, the role of\n                      <jats:italic toggle=\"yes\">msf<\/jats:italic>\n                      in other conditions and its molecular function remain unknown. To identify protein-protein interactions with Msf, a yeast two-hybrid screen against an\n                      <jats:italic toggle=\"yes\">H. influenzae<\/jats:italic>\n                      prey library was conducted, which found potential interactions with lipoprotein exporter protein LolD and an autotransporter adhesin Hap. To identify effects of\n                      <jats:italic toggle=\"yes\">msf<\/jats:italic>\n                      on gene expression, we compared wild-type and mutant strains grown in multiple culture conditions by RNA-seq. The results indicate that\n                      <jats:italic toggle=\"yes\">msf<\/jats:italic>\n                      modulates global gene expression in a condition-dependent manner, exerting an especially strong influence in starved surface-attached biofilm cells. The few consistent changes in mutants\u2019 planktonic exponential and stationary phases included decreased expression of two paralogous autotransporter adhesins. By contrast, mutant cells in starved surface-attached biofilms had dramatic changes in expression, including upregulation of protein translation and downregulation of alternative carbon metabolism. However, assays of 24 hour biofilm phenotypes found only subtle gene expression changes. Together, the results point to a speculative model of Msf functioning as an envelope-associated chaperone whose presence affects the relative expression of proteins at the outer membrane.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      Comparing genomes from different clinical isolates of the same pathogenic bacterial species has identified genes associated with virulence, but many of these are understudied or have no known function. The\n                      <jats:italic toggle=\"yes\">msf<\/jats:italic>\n                      gene was previously implicated as a virulence factor in\n                      <jats:italic toggle=\"yes\">Haemophilus influenzae<\/jats:italic>\n                      , a common cause of mucosal diseases including middle-ear and chronic lung infections. This study finds that the\n                      <jats:italic toggle=\"yes\">msf<\/jats:italic>\n                      gene causes condition-specific changes in gene expression, with especially dramatic changes in starved surface-attached biofilm cells. Along with identification of putative protein-protein interaction partners, the results provide new clues as to the molecular and cellular function of Msf, potentially as an envelope-associated chaperone involved in membrane protein trafficking. Understanding how virulence-associated genes like\n                      <jats:italic toggle=\"yes\">msf<\/jats:italic>\n                      modulate bacterial responses to the environment may help explain why some bacterial strains remain harmless colonizers while others become pathogens.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/mbio.01263-26","type":"journal-article","created":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T13:00:56Z","timestamp":1785330056000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":0,"title":["The\n                    <i>msf<\/i>\n                    gene causes condition-specific shifts in global gene expression in\n                    <i>Haemophilus influenzae<\/i>"],"prefix":"10.1128","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0009-0006-8853-2934","authenticated-orcid":true,"given":"Evangeline 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