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1544 (MDXLIV) was a leap year starting on Tuesday of the Julian calendar, the 1544th year of the Common Era (CE) and Anno Domini (AD) designations, the 544th year of the 2nd millennium, the 44th year of the 16th century, and the 5th year of the 1540s decade. As of the start of 1544, the Gregorian calendar was 10 days ahead of the Julian calendar, which was the dominant calendar of the time.
- January 13 – At Västerås, the estates of Sweden swear loyalty to King Gustav Vasa and to his heirs, ending the traditional electoral monarchy in Sweden. Gustav subsequently signs an alliance with the Kingdom of France.
- February 20 – The Fourth Diet of Speyer is convened.
- April 11 – Battle of Ceresole: French forces under the Comte d'Enghien defeat forces of the Holy Roman Empire, under the Marques Del Vasto, near Turin.
- May – Charles V, Holy Roman Emperor, again invades eastern France.
- May 3 – Edward Seymour, Earl of Hertford, with an English army, captures Leith and Edinburgh from the Kingdom of Scotland.
- June 19–August 18 – Troops of the Holy Roman Empire besiege Saint-Dizier, in eastern France.
- January 19 – King Francis II of France (d. 1560)
- January 24 – Gillis van Coninxloo, Flemish painter (d. 1607)
- February 3 – César de Bus, French Catholic priest (d. 1607)
- March 11 – Torquato Tasso, Italian poet (d. 1595)
- April – Thomas Fleming, English judge (d. 1613)
- April 20 – Renata of Lorraine, duchess consort of Bavaria (d. 1602)
- May 24 – William Gilbert, English scientist and astronomer (d. 1603)
- July 14 – Henry Compton, 1st Baron Compton, English politician (d. 1589)
- August 9 – Bogislaw XIII, Duke of Pomerania (d. 1606)
- September 1 – John Gordon, Scottish bishop (d. 1619)
- September 29 – Giovanni de' Medici, Italian Catholic cardinal (d. 1562)
- November 1 – Hasan Kafi Pruak, Bosnian scholar and judge (d. 1615)
- November 15 – Dorothea Susanne of Simmern, Duchess of Saxe-Weimar (d. 1592)
- December 23 – Anna of Saxony, only child and heiress of Maurice, Elector of Saxony (d. 1577)
- date unknown
- March 16 – Louis V, Elector Palatine (1508-1544) (b. 1478)
- March 22 – Johannes Magnus, last Catholic Archbishop of Sweden (b. 1488)
- April 30 – Thomas Audley, Lord Chancellor of England (b. 1488)
- June 14 – Antoine, Duke of Lorraine (b. 1489)
- July 15 – René of Châlon, Prince of the House of Orange (b. 1519)
- June 23 – Eleonore of Fürstenberg, wife of Philip IV, Count of Hanau-Lichtenberg (b. 1523)
- August 19 – Hans Buser, Swiss noble (b. 1513)
- September 12 – Clément Marot, French poet (b. 1496)
- September 25 – Valerius Cordus, German physician and scientist (b. 1515)
- October 10 – Charles Blount, 5th Baron Mountjoy, English courtier and patron of learning (b. 1516)
- October 12 – Antonio Pucci, Italian Catholic cardinal (b. 1485)
- November 13 – Ursula van Beckum, Dutch Anapabtist (b. 1520)
- November 15 – Lucy Brocadelli, Dominican tertiary and stigmatic (b. 1476)
- November 29 – Jungjong of Joseon (b. 1488)
- December 9 – Teofilo Folengo, Italian poet (b. 1491)
- date unknown